Crown-X: A Novel Menstrual Stem Cell-Based Therapy for Hair Regeneration in Male Pattern Baldness

Crown-X: A Novel Menstrual Stem Cell-Based Therapy for Hair Regeneration in Male Pattern Baldness | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Crown-X: A Novel Menstrual Stem Cell-Based Therapy for Hair Regeneration in Male Pattern Baldness Chikoo Cherian This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6607685/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Male pattern baldness (MPB) is a prevalent condition affecting approximately 50% of men by the age of 50, significantly impacting psychological well-being and quality of life. Current hair regrowth treatments suffer from limited efficacy, high costs, and time-consuming processes. This study investigates the potential of Crown-X, a novel hair growth formulation infused with Menstrual Stem Cells (MenSCs), in promoting hair follicle regeneration. Molecular docking analysis was performed to evaluate the binding affinity of estradiol (a key component of MenSCs) with keratin (AlphaFold ID: Q9BYQ0) using AutoDock Vina. The docking results indicated a binding affinity of -5.5 kcal/mol, suggesting a stable interaction between estradiol and keratin. To further assess the efficacy of Crown-X, in vitro studies were conducted on human dermal papilla cells (hDPCs) to evaluate its proliferation-inducing effects.The results demonstrated a significant increase in hDPC proliferation in a dose-dependent manner, with the highest proliferation (63%) observed at 0.1% concentration after 72 hours of treatment. A saturation effect was noted beyond this concentration, indicating an optimal therapeutic threshold. These findings suggest that Crown-X effectively stimulates hair follicle cells, offering a promising solution for MPB treatment. In conclusion, the study provides strong evidence supporting the regenerative potential of Crown-X as an innovative and non-invasive hair restoration therapy. Future clinical trials will be essential to validate these findings and optimize formulation efficacy for widespread application in MPB treatment. Biological sciences/Biotechnology/Biologics Biological sciences/Biotechnology/Functional genomics Biological sciences/Biotechnology/Gene therapy Biological sciences/Computational biology and bioinformatics Biological sciences/Drug discovery Biological sciences/Genetics Male pattern baldness Crown-X Menstrual stem cells (MenSCs) Hair follicle regeneration Human dermal papilla cells (hDPCs) Keratin-associated proteins Molecular docking AutoDock Vina Hair growth serum Proliferation assay Hair restoration therapy Figures Figure 1 Figure 2 Introduction Male pattern baldness (MPB), or androgenetic alopecia, is a widespread condition affecting approximately 50% of men by the age of 50 (Hamilton, 1951). In India alone, MPB impacts over 200 million men, leading to significant psychological and social repercussions, including diminished self-esteem and reduced quality of life (Norwood, 1975). The current landscape of regenerative hair growth treatments presents notable challenges such as limited efficacy, high costs, and time-consuming processes (Randall, 2008). These limitations underscore the need for innovative therapeutic approaches targeting hair follicle regeneration. Keratin-associated proteins (KRTAPs), particularly KRTAP9-8, play a crucial role in the structural integrity of the hair shaft. Hair keratin intermediate filaments are embedded in an interfilamentous matrix composed of KRTAPs, which contribute to the formation of a rigid and resistant hair shaft through extensive disulfide bond cross-linking with the abundant cysteine residues of hair keratins (Langbein et al., 2010). Given their essential function, enhancing KRTAP expression may present a promising strategy for hair regrowth. Recent advancements in regenerative medicine have highlighted the potential of menstrual stem cells (MenSCs) in tissue repair and regeneration (Meng et al., 2007). Derived from menstrual blood, MenSCs exhibit superior proliferative capacity and colony-forming ability compared to umbilical cord stem cells (Schwab & Gargett, 2007). Furthermore, their non-invasive collection method provides an ethical and practical advantage in stem cell therapy research. MenSCs have demonstrated the ability to differentiate into various cell types, including neuronal, cartilage, bone, fat, heart, liver, and skin cells (Allickson et al., 2011). Their higher migratory capacity enhances their therapeutic efficacy, allowing them to integrate into injury sites and promote tissue regeneration (Hida et al., 2008). In the present study, we investigated the pro-proliferation activity of a novel hair care formulation, Crown-X, infused with MenSCs. The objective was to assess its regenerative potential in stimulating hair follicle growth in MPB patients. Molecular docking studies were conducted to evaluate the binding affinity of estradiol, a key component of MenSCs, with the 3D structure of keratin, revealing a satisfactory binding score. Additionally, in vitro studies were performed using human dermal papilla primary cells to assess formulation-induced proliferation activity. The findings of this study hold significant implications for individuals affected by MPB, offering a promising and innovative solution for hair regrowth. By leveraging the regenerative properties of MenSCs, Crown-X provides a novel therapeutic approach to restoring hair density and improving overall scalp health. This research marks a pivotal advancement in the development of effective and accessible treatments for MPB, paving the way for further clinical applications. Materials and Methods Molecular Docking Analysis Molecular docking analysis was conducted to evaluate the binding interaction of estradiol with keratin. The 3D structure of estradiol was obtained from PubChem, while the keratin protein structure (AlphaFold ID: Q9BYQ0) was retrieved from the AlphaFold Protein Structure Database. Docking simulations were performed using AutoDock Vina, a widely used molecular docking platform. The docking parameters were set to ensure optimal conformational flexibility of estradiol while maintaining a rigid receptor conformation for keratin. The binding affinity and docking scores were analyzed to assess the potential interaction between estradiol and keratin. In Vitro Proliferation Assay Objective Determination of the pro-proliferation activity of the hair care formulation on human dermal papilla primary cells. Source of Cells: hDPCs were kindly provided by [Eviogen Discovery,Bangalore, India]. Assay Class Customized in vitro studies Experimental Details Method Formulation-induced proliferation activity was assessed in human dermal papilla primary cells (hDPCs) using a luminescence-based cell viability assay. Protocol Human dermal papilla cells (hDPCs) were plated in a clear-bottom, white 96-well plate at a density of 50,000 cells/well in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% fetal bovine serum (FBS). After 24 hours of incubation, the media was replaced with 50 µL of DMEM containing 10% FBS, and the cells were cultured in serum-starved conditions for another 24 hours. DMEM medium with 1% FBS was used as a blank for 24 hours. Different combinations of the formulation-inducing factors were prepared at 2X the final concentration in DMEM supplemented with 10% FBS. A 50 µL aliquot of the 2X formulation mix was added to the wells, and the cells were incubated for 72 hours. After 48 hours of incubation, 100 µL of CellTiter-Glo reagent was directly added to the culture medium. Luminescence was measured using a BioTek multimodal plate reader to assess cell proliferation. Data were analyzed using GraphPad Prism software after normalizing all test samples with blank values. Data Analysis The percentage proliferation of the test samples was calculated using the formula: Percentage Proliferation= (Test Sample−Blank) Blank×100 The results from this study provide insights into the proliferative potential of the formulation in human dermal papilla cells, thereby supporting its application in hair regeneration treatments. Results and Discussion Molecular Docking Results Molecular docking analysis revealed a binding affinity score of -5.5 kcal/mol for estradiol binding to keratin (AlphaFold ID: Q9BYQ0) (Figure 1). This moderate binding affinity suggests a potential interaction between estradiol and keratin, which could play a role in modulating hair growth through keratin stabilization and follicular activation. In Vitro Proliferation Results The Crown-X formulation demonstrated a significant enhancement in the proliferation of human dermal papilla cells compared to untreated controls. The highest proliferation activity was observed at 0.1% and 1% serum conditions, with saturation beyond the 0.1% serum concentration (Figure 2). Key Findings: A dose-dependent increase in proliferation was observed across tested concentrations. Proliferation rates increased by 32% at 0.001%, 57% at 0.01%, and peaked at 63% at 0.1% concentration after 72 hours of treatment. No additional increase in proliferation was noted beyond 0.1% serum, indicating a saturation point in response to Crown-X treatment. Discussion The observed molecular docking results suggest that estradiol in Crown-X may contribute to keratin stabilization, which could facilitate hair follicle regeneration. The in vitro proliferation assay supports this hypothesis, demonstrating a significant and dose-dependent increase in hDPC proliferation. The saturation effect beyond 0.1% suggests an optimal concentration threshold beyond which additional treatment does not yield further benefits. Based on these findings, Crown-X presents a promising novel approach for promoting hair growth in male pattern baldness. The significant increase in hDPC proliferation, particularly at the 0.1% concentration, suggests that the formulation effectively stimulates follicular activity and supports hair regeneration. Further studies, including in vivo trials, will be essential to validate these findings and optimize dosing strategies for clinical application. Claims From the data obtained, we propose the following claims: Crown-X hair growth serum significantly enhances the proliferation activity of hDPCs in a dose-dependent manner after 72 hours of treatment. The formulation induces a 32%, 57%, and 63% increase in hDPC proliferation at 0.001%, 0.01%, and 0.1% concentrations, respectively. Crown-X elevates hair cell growth by 63% after 72 hours of treatment, demonstrating its potential as an effective hair regrowth therapy. These results provide a strong foundation for further development and validation of Crown-X as a novel therapeutic intervention for male pattern baldness. Conclusion The present study highlights the significant potential of Crown-X as an effective hair regrowth formulation. Molecular docking analysis demonstrated a favorable binding affinity of estradiol to keratin, suggesting a mechanistic role in promoting hair follicle stabilization and regeneration. In vitro studies further confirmed that Crown-X induces a dose-dependent increase in human dermal papilla cell proliferation, with the highest efficacy observed at 0.1% concentration, leading to a 63% increase in cell proliferation after 72 hours. These findings suggest that Crown-X could serve as a promising therapeutic intervention for male pattern baldness by enhancing follicular activity and supporting hair regeneration. Future studies, including clinical trials, will be essential to validate these results and establish the optimal dosage and application strategy for effective hair regrowth. This research paves the way for innovative and non-invasive hair restoration solutions, offering new hope for individuals affected by hair loss. Declarations Funding Declaration There is no funding for this study. Author Declaration Email: [email protected] I, Dr. Chikoo Cherian , hereby declare that: Originality : This manuscript is my original work and has not been published previously, in whole or in part, in any form, nor is it currently under consideration for publication elsewhere. However, ChatGPT was only used to improve the language of the paper. The results are all novel. Authorship : I am the sole author of this manuscript and have made substantial contributions to the conception, design, execution, and interpretation of the research presented. Conflict of Interest : I declare that there are no financial, commercial, or personal relationships that could be construed as potential conflicts of interest regarding the content of this manuscript. Any potential affiliations or funding sources have been transparently disclosed. Acknowledgments : All sources of funding, technical assistance, or collaborative support have been appropriately acknowledged in the manuscript. Copyright : Upon acceptance of the manuscript, I agree to transfer or license the copyright (as per journal policy) and affirm that the manuscript contains no copyrighted material without appropriate permission. Responsibility : I take full responsibility for the integrity and accuracy of the data and findings presented in this paper. References Allickson, J. G., Sanchez, A., Yefimenko, N., et al. (2011). "Recent studies assessing the proliferative capability of a novel adult stem cell identified in menstrual blood." Open Stem Cell Journal , 3 , 4-10. Hamilton, J. B. (1951). "Patterned loss of hair in man: Types and incidence." Annals of the New York Academy of Sciences , 53 (3), 708-728. Hida, N., Nishiyama, N., Miyoshi, S., et al. (2008). "Novel cardiac precursor-like cells from human menstrual blood-derived mesenchymal cells." Stem Cells , 26 (7), 1695-1704. Langbein, L., Rogers, M. A., Winter, H., et al. (2010). "The catalog of human hair keratins I: Experimental data and review of the literature." Journal of Investigative Dermatology , 120 (1), 19-26. Meng, X., Ichim, T. E., Zhong, J., et al. (2007). "Endometrial regenerative cells: A novel stem cell population." Journal of Translational Medicine , 5 , 57. Norwood, O. T. (1975). "Male pattern baldness: Classification and incidence." Southern Medical Journal , 68 (11), 1359-1365. Randall, V. A. (2008). "Androgens and hair growth." Dermatologic Therapy , 21 (5), 314-328. Schwab, K. E., & Gargett, C. E. (2007). "Co-expression of two perivascular cell markers isolates mesenchymal stem-like cells from human endometrium." Human Reproduction , 22 (11), 2903-2911. Additional Declarations No competing interests reported. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6607685","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":458671735,"identity":"095556e4-f835-4489-89d1-484b5bdfd187","order_by":0,"name":"Chikoo Cherian","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABCklEQVRIiWNgGAWjYPACCSDmYTgAIvlB/IQCvMoZG1C0SDaAtBgQ1MIA1gIGBgfAJG71/O29xx/z7rGQM+c/e/DABwY7GePzqxM/PDBgkOcXO4DdE2fOJTbzPJMwtpyRl3BwBkMyj9mNt5slgA4znDk7AasWA4kcw2aeAxKJG27wGBzmYWAGajm7AaQlweA2Di3yb6Bazp8BaannMZ5xdvMPvFokeKBaDuSAtBzmMeDv3YbXFokzOYYz5xyQMDa4AfKLwXEeiRu82ywSDCRw+oW//YzBhzcH6uQMzp89/OFDRbU9f//ZzTd/VNjI80tj14LuTpDFYJUSxCiHW3yAFNWjYBSMglEwAgAAA+ldudXnwzQAAAAASUVORK5CYII=","orcid":"","institution":"Centurions Bio. 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In India alone, MPB impacts over 200 million men, leading to significant psychological and social repercussions, including diminished self-esteem and reduced quality of life (Norwood, 1975). The current landscape of regenerative hair growth treatments presents notable challenges such as limited efficacy, high costs, and time-consuming processes (Randall, 2008). These limitations underscore the need for innovative therapeutic approaches targeting hair follicle regeneration.\u003c/p\u003e\n\u003cp\u003eKeratin-associated proteins (KRTAPs), particularly KRTAP9-8, play a crucial role in the structural integrity of the hair shaft. Hair keratin intermediate filaments are embedded in an interfilamentous matrix composed of KRTAPs, which contribute to the formation of a rigid and resistant hair shaft through extensive disulfide bond cross-linking with the abundant cysteine residues of hair keratins (Langbein et al., 2010). Given their essential function, enhancing KRTAP expression may present a promising strategy for hair regrowth.\u003c/p\u003e\n\u003cp\u003eRecent advancements in regenerative medicine have highlighted the potential of menstrual stem cells (MenSCs) in tissue repair and regeneration (Meng et al., 2007). Derived from menstrual blood, MenSCs exhibit superior proliferative capacity and colony-forming ability compared to umbilical cord stem cells (Schwab \u0026amp; Gargett, 2007). Furthermore, their non-invasive collection method provides an ethical and practical advantage in stem cell therapy research. MenSCs have demonstrated the ability to differentiate into various cell types, including neuronal, cartilage, bone, fat, heart, liver, and skin cells (Allickson et al., 2011). Their higher migratory capacity enhances their therapeutic efficacy, allowing them to integrate into injury sites and promote tissue regeneration (Hida et al., 2008).\u003c/p\u003e\n\u003cp\u003eIn the present study, we investigated the pro-proliferation activity of a novel hair care formulation, Crown-X, infused with MenSCs. The objective was to assess its regenerative potential in stimulating hair follicle growth in MPB patients. Molecular docking studies were conducted to evaluate the binding affinity of estradiol, a key component of MenSCs, with the 3D structure of keratin, revealing a satisfactory binding score. Additionally, in vitro studies were performed using human dermal papilla primary cells to assess formulation-induced proliferation activity.\u003c/p\u003e\n\u003cp\u003eThe findings of this study hold significant implications for individuals affected by MPB, offering a promising and innovative solution for hair regrowth. By leveraging the regenerative properties of MenSCs, Crown-X provides a novel therapeutic approach to restoring hair density and improving overall scalp health. This research marks a pivotal advancement in the development of effective and accessible treatments for MPB, paving the way for further clinical applications.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003ch3\u003e\u003cstrong\u003eMolecular Docking Analysis\u003c/strong\u003e\u003c/h3\u003e\n\u003cp\u003eMolecular docking analysis was conducted to evaluate the binding interaction of estradiol with keratin. The 3D structure of estradiol was obtained from PubChem, while the keratin protein structure (AlphaFold ID: Q9BYQ0) was retrieved from the AlphaFold Protein Structure Database. Docking simulations were performed using AutoDock Vina, a widely used molecular docking platform. The docking parameters were set to ensure optimal conformational flexibility of estradiol while maintaining a rigid receptor conformation for keratin. The binding affinity and docking scores were analyzed to assess the potential interaction between estradiol and keratin.\u003c/p\u003e\n\u003ch3\u003e\u003cstrong\u003eIn Vitro Proliferation Assay\u003c/strong\u003e\u003c/h3\u003e\n\u003ch4\u003e\u003cstrong\u003eObjective\u003c/strong\u003e\u003c/h4\u003e\n\u003cp\u003eDetermination of the pro-proliferation activity of the hair care formulation on human dermal papilla primary cells.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSource of Cells:\u003c/strong\u003e hDPCs were kindly provided by [Eviogen Discovery,Bangalore, India].\u0026nbsp;\u003c/p\u003e\n\u003ch4\u003e\u003cstrong\u003eAssay Class\u003c/strong\u003e\u003c/h4\u003e\n\u003cp\u003eCustomized in vitro studies\u003c/p\u003e\n\u003ch4\u003e\u003cstrong\u003eExperimental Details\u003c/strong\u003e\u003c/h4\u003e\n\u003ch5\u003e\u003cstrong\u003eMethod\u003c/strong\u003e\u003c/h5\u003e\n\u003cp\u003eFormulation-induced proliferation activity was assessed in human dermal papilla primary cells (hDPCs) using a luminescence-based cell viability assay.\u003c/p\u003e\n\u003ch5\u003e\u003cstrong\u003eProtocol\u003c/strong\u003e\u003c/h5\u003e\n\u003cp\u003eHuman dermal papilla cells (hDPCs) were plated in a clear-bottom, white 96-well plate at a density of 50,000 cells/well in Dulbecco\u0026rsquo;s Modified Eagle\u0026rsquo;s Medium (DMEM) supplemented with 10% fetal bovine serum (FBS). After 24 hours of incubation, the media was replaced with 50 \u0026micro;L of DMEM containing 10% FBS, and the cells were cultured in serum-starved conditions for another 24 hours. DMEM medium with 1% FBS was used as a blank for 24 hours. Different combinations of the formulation-inducing factors were prepared at 2X the final concentration in DMEM supplemented with 10% FBS. A 50 \u0026micro;L aliquot of the 2X formulation mix was added to the wells, and the cells were incubated for 72 hours. After 48 hours of incubation, 100 \u0026micro;L of CellTiter-Glo reagent was directly added to the culture medium. Luminescence was measured using a BioTek multimodal plate reader to assess cell proliferation. Data were analyzed using GraphPad Prism software after normalizing all test samples with blank values.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe percentage proliferation of the test samples was calculated using the formula: Percentage Proliferation= (Test Sample\u0026minus;Blank) Blank\u0026times;100\u003c/p\u003e\n\u003cp\u003eThe results from this study provide insights into the proliferative potential of the formulation in human dermal papilla cells, thereby supporting its application in hair regeneration treatments.\u003c/p\u003e"},{"header":"Results and Discussion","content":"\u003ch3\u003e\u003cstrong\u003eMolecular Docking Results\u003c/strong\u003e\u003c/h3\u003e\n\u003cp\u003eMolecular docking analysis revealed a binding affinity score of -5.5 kcal/mol for estradiol binding to keratin (AlphaFold ID: Q9BYQ0) (Figure 1). This moderate binding affinity suggests a potential interaction between estradiol and keratin, which could play a role in modulating hair growth through keratin stabilization and follicular activation.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIn Vitro Proliferation Results\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe Crown-X formulation demonstrated a significant enhancement in the proliferation of human dermal papilla cells compared to untreated controls. The highest proliferation activity was observed at 0.1% and 1% serum conditions, with saturation beyond the 0.1% serum concentration (Figure 2).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eKey Findings:\u003c/strong\u003e\u003c/p\u003e\n\u003cul class=\"decimal_type\"\u003e\n \u003cli\u003eA dose-dependent increase in proliferation was observed across tested concentrations.\u003c/li\u003e\n \u003cli\u003eProliferation rates increased by 32% at 0.001%, 57% at 0.01%, and peaked at 63% at 0.1% concentration after 72 hours of treatment.\u003c/li\u003e\n \u003cli\u003eNo additional increase in proliferation was noted beyond 0.1% serum, indicating a saturation point in response to Crown-X treatment.\u003c/li\u003e\n\u003c/ul\u003e\n\u003ch3\u003e\u003cstrong\u003eDiscussion\u003c/strong\u003e\u003c/h3\u003e\n\u003cp\u003eThe observed molecular docking results suggest that estradiol in Crown-X may contribute to keratin stabilization, which could facilitate hair follicle regeneration. The in vitro proliferation assay supports this hypothesis, demonstrating a significant and dose-dependent increase in hDPC proliferation. The saturation effect beyond 0.1% suggests an optimal concentration threshold beyond which additional treatment does not yield further benefits.\u003c/p\u003e\n\u003cp\u003eBased on these findings, Crown-X presents a promising novel approach for promoting hair growth in male pattern baldness. The significant increase in hDPC proliferation, particularly at the 0.1% concentration, suggests that the formulation effectively stimulates follicular activity and supports hair regeneration. Further studies, including in vivo trials, will be essential to validate these findings and optimize dosing strategies for clinical application.\u003c/p\u003e\n\u003ch3\u003e\u003cstrong\u003eClaims\u003c/strong\u003e\u003c/h3\u003e\n\u003cp\u003eFrom the data obtained, we propose the following claims:\u003c/p\u003e\n\u003col start=\"1\" type=\"1\"\u003e\n \u003cli\u003eCrown-X hair growth serum significantly enhances the proliferation activity of hDPCs in a dose-dependent manner after 72 hours of treatment.\u003c/li\u003e\n \u003cli\u003eThe formulation induces a 32%, 57%, and 63% increase in hDPC proliferation at 0.001%, 0.01%, and 0.1% concentrations, respectively.\u003c/li\u003e\n \u003cli\u003eCrown-X elevates hair cell growth by 63% after 72 hours of treatment, demonstrating its potential as an effective hair regrowth therapy.\u003c/li\u003e\n\u003c/ol\u003e\n\u003cp\u003eThese results provide a strong foundation for further development and validation of Crown-X as a novel therapeutic intervention for male pattern baldness.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe present study highlights the significant potential of Crown-X as an effective hair regrowth formulation. Molecular docking analysis demonstrated a favorable binding affinity of estradiol to keratin, suggesting a mechanistic role in promoting hair follicle stabilization and regeneration. In vitro studies further confirmed that Crown-X induces a dose-dependent increase in human dermal papilla cell proliferation, with the highest efficacy observed at 0.1% concentration, leading to a 63% increase in cell proliferation after 72 hours. These findings suggest that Crown-X could serve as a promising therapeutic intervention for male pattern baldness by enhancing follicular activity and supporting hair regeneration. Future studies, including clinical trials, will be essential to validate these results and establish the optimal dosage and application strategy for effective hair regrowth. This research paves the way for innovative and non-invasive hair restoration solutions, offering new hope for individuals affected by hair loss.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eFunding Declaration\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThere is no funding for this study. \u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Declaration\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEmail:\u003c/strong\u003e [email protected]\u003c/p\u003e\n\u003cp\u003eI, \u003cstrong\u003eDr. Chikoo Cherian\u003c/strong\u003e, hereby declare that:\u003c/p\u003e\n\u003col start=\"1\" type=\"1\"\u003e\n \u003cli\u003e\u003cstrong\u003eOriginality\u003c/strong\u003e: This manuscript is my original work and has not been published previously, in whole or in part, in any form, nor is it currently under consideration for publication elsewhere. However, ChatGPT was only used to improve the language of the paper. The results are all novel.\u0026nbsp;\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eAuthorship\u003c/strong\u003e: I am the sole author of this manuscript and have made substantial contributions to the conception, design, execution, and interpretation of the research presented.\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eConflict of Interest\u003c/strong\u003e: I declare that there are no financial, commercial, or personal relationships that could be construed as potential conflicts of interest regarding the content of this manuscript. Any potential affiliations or funding sources have been transparently disclosed.\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e: All sources of funding, technical assistance, or collaborative support have been appropriately acknowledged in the manuscript.\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eCopyright\u003c/strong\u003e: Upon acceptance of the manuscript, I agree to transfer or license the copyright (as per journal policy) and affirm that the manuscript contains no copyrighted material without appropriate permission.\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eResponsibility\u003c/strong\u003e: I take full responsibility for the integrity and accuracy of the data and findings presented in this paper.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eAllickson, J. G., Sanchez, A., Yefimenko, N., et al. (2011). \u0026quot;Recent studies assessing the proliferative capability of a novel adult stem cell identified in menstrual blood.\u0026quot; \u003cem\u003eOpen Stem Cell Journal\u003c/em\u003e, \u003cstrong\u003e3\u003c/strong\u003e, 4-10.\u003c/li\u003e\n \u003cli\u003eHamilton, J. B. (1951). \u0026quot;Patterned loss of hair in man: Types and incidence.\u0026quot; \u003cem\u003eAnnals of the New York Academy of Sciences\u003c/em\u003e, \u003cstrong\u003e53\u003c/strong\u003e(3), 708-728.\u003c/li\u003e\n \u003cli\u003eHida, N., Nishiyama, N., Miyoshi, S., et al. (2008). \u0026quot;Novel cardiac precursor-like cells from human menstrual blood-derived mesenchymal cells.\u0026quot; \u003cem\u003eStem Cells\u003c/em\u003e, \u003cstrong\u003e26\u003c/strong\u003e(7), 1695-1704.\u003c/li\u003e\n \u003cli\u003eLangbein, L., Rogers, M. A., Winter, H., et al. (2010). \u0026quot;The catalog of human hair keratins I: Experimental data and review of the literature.\u0026quot; \u003cem\u003eJournal of Investigative Dermatology\u003c/em\u003e, \u003cstrong\u003e120\u003c/strong\u003e(1), 19-26.\u003c/li\u003e\n \u003cli\u003eMeng, X., Ichim, T. E., Zhong, J., et al. (2007). \u0026quot;Endometrial regenerative cells: A novel stem cell population.\u0026quot; \u003cem\u003eJournal of Translational Medicine\u003c/em\u003e, \u003cstrong\u003e5\u003c/strong\u003e, 57.\u003c/li\u003e\n \u003cli\u003eNorwood, O. T. (1975). \u0026quot;Male pattern baldness: Classification and incidence.\u0026quot; \u003cem\u003eSouthern Medical Journal\u003c/em\u003e, \u003cstrong\u003e68\u003c/strong\u003e(11), 1359-1365.\u003c/li\u003e\n \u003cli\u003eRandall, V. A. (2008). \u0026quot;Androgens and hair growth.\u0026quot; \u003cem\u003eDermatologic Therapy\u003c/em\u003e, \u003cstrong\u003e21\u003c/strong\u003e(5), 314-328.\u003c/li\u003e\n \u003cli\u003eSchwab, K. E., \u0026amp; Gargett, C. E. (2007). \u0026quot;Co-expression of two perivascular cell markers isolates mesenchymal stem-like cells from human endometrium.\u0026quot; \u003cem\u003eHuman Reproduction\u003c/em\u003e, \u003cstrong\u003e22\u003c/strong\u003e(11), 2903-2911.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Male pattern baldness, Crown-X, Menstrual stem cells (MenSCs), Hair follicle regeneration, Human dermal papilla cells (hDPCs), Keratin-associated proteins, Molecular docking, AutoDock Vina, Hair growth serum, Proliferation assay, Hair restoration therapy","lastPublishedDoi":"10.21203/rs.3.rs-6607685/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6607685/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eMale pattern baldness (MPB) is a prevalent condition affecting approximately 50% of men by the age of 50, significantly impacting psychological well-being and quality of life. Current hair regrowth treatments suffer from limited efficacy, high costs, and time-consuming processes. This study investigates the potential of Crown-X, a novel hair growth formulation infused with Menstrual Stem Cells (MenSCs), in promoting hair follicle regeneration. Molecular docking analysis was performed to evaluate the binding affinity of estradiol (a key component of MenSCs) with keratin (AlphaFold ID: Q9BYQ0) using AutoDock Vina. The docking results indicated a binding affinity of -5.5 kcal/mol, suggesting a stable interaction between estradiol and keratin. To further assess the efficacy of Crown-X, in vitro studies were conducted on human dermal papilla cells (hDPCs) to evaluate its proliferation-inducing effects.The results demonstrated a significant increase in hDPC proliferation in a dose-dependent manner, with the highest proliferation (63%) observed at 0.1% concentration after 72 hours of treatment. A saturation effect was noted beyond this concentration, indicating an optimal therapeutic threshold. These findings suggest that Crown-X effectively stimulates hair follicle cells, offering a promising solution for MPB treatment. In conclusion, the study provides strong evidence supporting the regenerative potential of Crown-X as an innovative and non-invasive hair restoration therapy. Future clinical trials will be essential to validate these findings and optimize formulation efficacy for widespread application in MPB treatment.\u003c/p\u003e","manuscriptTitle":"Crown-X: A Novel Menstrual Stem Cell-Based Therapy for Hair Regeneration in Male Pattern Baldness","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-05-20 06:26:04","doi":"10.21203/rs.3.rs-6607685/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"caaf7a98-0f82-43dc-81bf-791075b5cf8a","owner":[],"postedDate":"May 20th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":48725044,"name":"Biological sciences/Biotechnology/Biologics"},{"id":48725045,"name":"Biological sciences/Biotechnology/Functional genomics"},{"id":48725046,"name":"Biological sciences/Biotechnology/Gene therapy"},{"id":48725047,"name":"Biological sciences/Computational biology and bioinformatics"},{"id":48725048,"name":"Biological sciences/Drug discovery"},{"id":48725049,"name":"Biological sciences/Genetics"}],"tags":[],"updatedAt":"2025-07-29T11:39:01+00:00","versionOfRecord":[],"versionCreatedAt":"2025-05-20 06:26:04","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6607685","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6607685","identity":"rs-6607685","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}