Exploring the Potential of Aloe Vera Hydro-Alcoholic Leaf Extract for Topical Diabetic Wound Healing Treatment
Abstract
Background: Diabetic wounds present substantial challenges within the diabetic patient population, prompting diverse therapeutic strategies including the exploration of natural remedies. Methods: This investigation aims to comprehensively evaluate the efficacy of topical application of Aloe vera hydro-alcoholic leaf extract (AVHE) in addressing the various contributing elements to delayed wound healing in the diabetes through cell based assays and animal model. Results: The AVHE exhibited remarkable α-amylase inhibitory activity, indicated by an IC50 value of 0.051 mg/mL, comparable to the standard Acarbose (IC50 = 0.041 mg/mL). Fluorescent microscopy analysis demonstrated significant glucose uptake by McCoy fibroblast cells treated with AVHE, evidenced by 187.74% uptake of 2-NBDG at concentration of 0.1 mg/mL. Furthermore, the AVHE exhibited notable anti-bacterial efficacy against a spectrum of tested pathogens, with pronounced effectiveness against Bacillus subtilis and Staphylococcus aureus. The AVHE displayed enhanced wound closure, achieving a remarkable 57.03 % closure rate at concentration of 0.1 mg/mL compared to the untreated cells 10.01% in an in vitro scratch assay conducted in McCoy cells. In vivo assessments included evaluating the potential of AVHE to induce caudal fin regeneration in Zebra fish, revealing a significant 60% development within 7 days post-amputation. Conclusion: Collectively, these findings highlight the therapeutic potential of topical application of AVHE for the advancement of treatments targeting diabetic foot ulcers.
Keywords:
Diabetic wounds; Aloe vera; Anti-diabetic; In vivo.
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References
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Babu, S. N., Govindarajan, S., Vijayalakshmi, M. A., & Noor, A. (2019). Evaluation of in vitro anti-diabetic and anti-oxidant activities and preliminary phytochemical screening of gel, epidermis and flower extract of Aloe vera. Research journal of pharmacy and technology, 12(4), 1761-1768. http://dx.doi.org/10.5958/0974-360X.2019.00295.6
Berbudi, A., Khairani, S., Tjahjadi, AI. (2025). Interplay Between Insulin Resistance and Immune Dysregulation in Type 2 Diabetes Mellitus: Implications for Therapeutic Interventions. Immunotargets Ther, 14, 359-382. https://doi.org/10.2147/ITT.S499605.
Bhardwaj, K., & Dubey, W. (2019). Quantitative analysis of primary and secondary metabolites of ethanol seed extract of Origanum majorana (Marjoram). Journal of Pharmacognosy and Phytochemistry, 8(1), 1251-1255. Retrieved from: https://www.phytojournal.com/archives/2019/vol8issue1/PartU/7-6-551-580.pdf. Accessed on 19th August, 2024.
Blois, M. S. (1958). Antioxidant determinations by the use of a stable free radical. Nature, 181, 1199-1200. https://doi.org/10.1038/1811199a0
Catalano, A., Ceramella, J., Iacopetta, D., Marra, M., Conforti, F., Lupi, F. R., Gabriele, D., Borges, F., & Sinicropi, M. S. (2024). Aloe vera-An Extensive Review Focused on Recent Studies. Foods (Basel, Switzerland), 13(13). https://doi.org/10.3390/foods13132155
Chelu, M., Calderon Moreno, J. M., Musuc, A. M., & Popa, M. (2024). Natural Regenerative Hydrogels for Wound Healing. Gels (Basel, Switzerland), 10(9), 547. https://doi.org/10.3390/gels10090547
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Chelu, M., Musuc, A. M., Popa, M., & Calderon Moreno, J. (2023b). Aloe vera-Based Hydrogels for Wound Healing: Properties and Therapeutic Effects. Gels (Basel, Switzerland), 9(7), 539. https://doi.org/10.3390/gels9070539
Chen, X., Xie, N., Feng, L., Huang, Y., Wu, Y., Zhu, H., ... & Zhang, Y. (2025). Oxidative stress in diabetes mellitus and its complications: From pathophysiology to therapeutic strategies. Chinese Medical Journal, 138(1), 15-27. https://doi.org/10.1097/CM9.0000000000003230
CLSI (2012) Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grows Aerobically. Approved Standard, 9th Edition, CLSI Document MO7- A9, Clinical and Laboratory Standards Institute, Wayne.P.A.
El Hosry, L., Sok, N., Richa, R., Al Mashtoub, L., Cayot, P., & Bou-Maroun, E. (2023). Sample preparation and analytical techniques in the determination of trace elements in food: A review. Foods, 12(4), 895. https://doi.org/10.3390/foods12040895
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Girgin, R., Çeker, G., Turan, İ., Kutu, F., Elma, Y., & Can, E. Y. (2024). The effects of topical aloe vera on an experimentally designed penile fracture model in rats. Turkish Journal of Trauma & Emergency Surgery, 30(3), 147. https://doi.org/10.14744/tjtes.2024.26425
Hajfathalian, M., Ghelichi, S., & Jacobsen, C. (2025). Anti‐obesity peptides from food: Production, evaluation, sources, and commercialization. Comprehensive Reviews in Food Science and Food Safety, 24(2). https://doi.org/10.1111/1541-4337.70158
Hedge, J. E., Hofreiter, B. T., & Whistler, R. L. (1962). Carbohydrate chemistry. Academic Press, New York, 17, 371-380.
Isopencu, G. O., Covaliu-Mierlă, C. I., & Deleanu, I. M. (2023). From Plants to Wound Dressing and Transdermal Delivery of Bioactive Compounds. Plants (Basel, Switzerland), 12(14). https://doi.org/10.3390/plants12142661
Kaczmarek-Szczepańska, B., Glajc, P., Chmielniak, D., Gwizdalska, K., Swiontek Brzezinska, M., Dembinska, K., ... & Zasada, L. (2025). Development and Characterization of Biocompatible Chitosan-Aloe Vera Films Functionalized with Gluconolactone and Sorbitol for Advanced Wound Healing Applications. ACS Applied Materials & Interfaces, 17(10). https://doi.org/10.1021/acsami.5c00715
Liang, J., Cui, L., Li, J., Guan, S., Zhang, K., & Li, J. (2021). Aloe vera: a medicinal plant used in skin wound healing. Tissue Engineering Part B: Reviews, 27(5), 455-474. https://doi.org/10.1089/ten.teb.2020.0236
Lowry, O. H., Rosebrough, N. J., Farr, A. L., & Randall, R. J. (1951). Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry, 193(1), 265-275. https://doi.org/10.1016/S0021-9258(19)52451-6
Mahesh, G., Anil Kumar, K., & Reddanna, P. (2021). Overview on the discovery and development of anti-inflammatory drugs: should the focus be on synthesis or degradation of PGE2?. Journal of Inflammation Research, 253-263. https://doi.org/10.2147/JIR.S278514
Mohsin, F., Javaid, S., Tariq, M., & Mustafa, M. (2024). Molecular immunological mechanisms of impaired wound healing in diabetic foot ulcers (DFU), current therapeutic strategies and future directions. International Immunopharmacology, 139. https://doi.org/10.1016/j.intimp.2024.112713
Muller, M. J., Hollyoak, M. A., Moaveni, Z., Brown, T. L. H., Herndon, D. N., & Heggers, J. P. (2003). Retardation of wound healing by silver sulfadiazine is reversed by Aloe vera and nystatin. Burns, 29(8), 834-836. https://doi.org/10.1016/s0305-4179(03)00198-0
Negahdari, S., Galehdari, H., Kesmati, M., Rezaie, A., & Shariati, G. (2017). Wound healing activity of extracts and formulations of aloe vera, henna, adiantum capillus-veneris, and myrrh on mouse dermal fibroblast cells. International Journal of Preventive Medicine, 8(1), 18. https://doi.org/10.4103/ijpvm.IJPVM_338_16
Okur, M. E., Karadağ, A. E., Özhan, Y., Sipahi, H., Ayla, Ş., Daylan, B., ... & Demirci, F. (2021). Anti-inflammatory, analgesic and in vivo-in vitro wound healing potential of the Phlomis rigida Labill. extract. Journal of Ethnopharmacology, 266. https://doi.org/10.1016/j.jep.2020.113408
Patitucci, F., Motta, M. F., Mileti, O., Dattilo, M., Malivindi, R., Pezzi, G., ... & Puoci, F. (2025). Enhancing burn wound care with pre-crosslinked 3D-printed patches: Bromelain delivery and aloe vera bioactives integration for improved healing outcomes. International Journal of Pharmaceutics. https://doi.org/10.1016/j.ijpharm.2025.125304
Rasband, W.S. ImageJ, U. S. (1997). National Institutes of Health, Bethesda, Maryland, USA, 1997-2018. https://imagej.nih.gov/ij/
Re, R., Pellegrini, N., Proteggente, A., Pannala, A., Yang, M., & Rice-Evans, C. (1999). Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biology and Medicine, 26(9-10), 1231-1237. https://doi.org/10.1016/s0891-5849(98)00315-3
Richardson, R., Slanchev, K., Kraus, C., Knyphausen, P., Eming, S., & Hammerschmidt, M. (2013). Adult zebrafish as a model system for cutaneous wound-healing research. Journal of Investigative Dermatology, 133(6), 1655-1665. https://doi.org/10.1038/jid.2013.16
Saeed, M. K., Zahra, N., Abidi, S. H. I., & Syed, Q. U. A. (2022). Phytochemical screening and DPPH free radical scavenging activity of Aloe vera (Aloe barbadensis Miller) powder. International Journal of Food Science and Agriculture, 6(3). http://dx.doi.org/10.26855/ijfsa.2022.09.010
Sena, C. L., Konig, I., da Silva, M. H., Carapiá, M. S., Braga, M. A., & Marcussi, S. (2025). Effects of Uncaria tomentosa and Uncaria guianensis aqueous extract on enzyme activity and modulation of hemostasis. Scientia Plena, 21(2). https://doi.org/10.14808/sci.plena.2025.021101
Singleton, V. L., Orthofer, R., & Lamuela-Raventós, R. M. (1999). [14] Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. In Methods in Enzymology. 299, 152-178.Academic press. https://doi.org/10.1016/S0076-6879(99)99017-1
Smith, J., & Rai, V. (2024). Novel factors regulating proliferation, migration, and differentiation of fibroblasts, keratinocytes, and vascular smooth muscle cells during wound healing. Biomedicines, 12(9). https://doi.org/10.3390/biomedicines12091939
Sumner, J. B., & Graham, V. A. (1921). Dinitrosalicylic acid: a reagent for the estimation of sugar in normal and diabetic urine. Journal of Biological Chemistry, 47(1), 5-9. https://doi.org/10.1016/S0021-9258(18)86093-8
Tomic, D., Shaw, J. E., & Magliano, D. J. (2022). The burden and risks of emerging complications of diabetes mellitus. Nature Reviews Endocrinology, 18(9), 525-539. https://doi.org/10.1038/s41574-022-00690-7
Wang, X., Yang, J., Zhao, Q., Xie, X., Deng, F., Wang, Z., ... & Lv, D. (2024). A tissue-adhesive, mechanically enhanced, natural Aloe Vera-based injectable hydrogel for wound healing: Macrophage mediation and collagen proliferation. International Journal of Biological Macromolecules, 283. https://doi.org/10.1016/j.ijbiomac.2024.137452
Wilkinson, H. N., & Hardman, M. J. (2020). Wound healing: cellular mechanisms and pathological outcomes. Open Biology, 10(9). https://doi.org/10.1098/rsob.200223
Woisky, R. G., & Salatino, A. (1998). Analysis of propolis: some parameters and procedures for chemical quality control. Journal of Apicultural Research, 37(2), 99-105. https://doi.org/10.1080/00218839.1998.11100961
World Health Organization (WHO). (1998). Quality control methods for medicinal plant materials. World Health Organization. Retrieved from: https://www.who.int/docs/default-source/medicines/norms-and-standards/guidelines/quality-control/quality-control-methods-for-medicinal-plant-materials.pdf?sfvrsn=b451e7c6_0. Accessed on 10th August, 2024.
Yousif, D., Yousif, Z., & Joseph, P. (2024). Diabetic Foot Ulcer Neuropathy, Impaired Vasculature, and Immune Responses. In Diabetic Foot Ulcers-Pathogenesis, Innovative Treatments and AI Applications. IntechOpen. https://doi.org/10.5772/intechopen.1003834
Arafa, N. M. S., Hummadi, H. M. A., & Badr, G. M. (2025). The potential of aloe vera gel utilization for skin wound healing in rats based on GC–MS and HPLC chemical profile. The Journal of Basic and Applied Zoology, 86(1), 6.https://doi.org/10.1186/s41936-024-00424-3
Babu, S. N., Govindarajan, S., Vijayalakshmi, M. A., & Noor, A. (2019). Evaluation of in vitro anti-diabetic and anti-oxidant activities and preliminary phytochemical screening of gel, epidermis and flower extract of Aloe vera. Research journal of pharmacy and technology, 12(4), 1761-1768. http://dx.doi.org/10.5958/0974-360X.2019.00295.6
Berbudi, A., Khairani, S., Tjahjadi, AI. (2025). Interplay Between Insulin Resistance and Immune Dysregulation in Type 2 Diabetes Mellitus: Implications for Therapeutic Interventions. Immunotargets Ther, 14, 359-382. https://doi.org/10.2147/ITT.S499605.
Bhardwaj, K., & Dubey, W. (2019). Quantitative analysis of primary and secondary metabolites of ethanol seed extract of Origanum majorana (Marjoram). Journal of Pharmacognosy and Phytochemistry, 8(1), 1251-1255. Retrieved from: https://www.phytojournal.com/archives/2019/vol8issue1/PartU/7-6-551-580.pdf. Accessed on 19th August, 2024.
Blois, M. S. (1958). Antioxidant determinations by the use of a stable free radical. Nature, 181, 1199-1200. https://doi.org/10.1038/1811199a0
Catalano, A., Ceramella, J., Iacopetta, D., Marra, M., Conforti, F., Lupi, F. R., Gabriele, D., Borges, F., & Sinicropi, M. S. (2024). Aloe vera-An Extensive Review Focused on Recent Studies. Foods (Basel, Switzerland), 13(13). https://doi.org/10.3390/foods13132155
Chelu, M., Calderon Moreno, J. M., Musuc, A. M., & Popa, M. (2024). Natural Regenerative Hydrogels for Wound Healing. Gels (Basel, Switzerland), 10(9), 547. https://doi.org/10.3390/gels10090547
Chelu, M., Musuc, A. M., Aricov, L., Ozon, E. A., Iosageanu, A., Stefan, L. M., Prelipcean, A. M., Popa, M., & Moreno, J. C. (2023a). Antibacterial Aloe vera Based Biocompatible Hydrogel for Use in Dermatological Applications. International Journal of Molecular Sciences, 24(4), 3893. https://doi.org/10.3390/ijms24043893
Chelu, M., Musuc, A. M., Popa, M., & Calderon Moreno, J. (2023b). Aloe vera-Based Hydrogels for Wound Healing: Properties and Therapeutic Effects. Gels (Basel, Switzerland), 9(7), 539. https://doi.org/10.3390/gels9070539
Chen, X., Xie, N., Feng, L., Huang, Y., Wu, Y., Zhu, H., ... & Zhang, Y. (2025). Oxidative stress in diabetes mellitus and its complications: From pathophysiology to therapeutic strategies. Chinese Medical Journal, 138(1), 15-27. https://doi.org/10.1097/CM9.0000000000003230
CLSI (2012) Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grows Aerobically. Approved Standard, 9th Edition, CLSI Document MO7- A9, Clinical and Laboratory Standards Institute, Wayne.P.A.
El Hosry, L., Sok, N., Richa, R., Al Mashtoub, L., Cayot, P., & Bou-Maroun, E. (2023). Sample preparation and analytical techniques in the determination of trace elements in food: A review. Foods, 12(4), 895. https://doi.org/10.3390/foods12040895
Floris, B., Galloni, P., Conte, V., & Sabuzi, F. (2021). Tailored functionalization of natural phenols to improve biological activity. Biomolecules, 11(9). https://doi.org/10.3390/biom11091325
Girgin, R., Çeker, G., Turan, İ., Kutu, F., Elma, Y., & Can, E. Y. (2024). The effects of topical aloe vera on an experimentally designed penile fracture model in rats. Turkish Journal of Trauma & Emergency Surgery, 30(3), 147. https://doi.org/10.14744/tjtes.2024.26425
Hajfathalian, M., Ghelichi, S., & Jacobsen, C. (2025). Anti‐obesity peptides from food: Production, evaluation, sources, and commercialization. Comprehensive Reviews in Food Science and Food Safety, 24(2). https://doi.org/10.1111/1541-4337.70158
Hedge, J. E., Hofreiter, B. T., & Whistler, R. L. (1962). Carbohydrate chemistry. Academic Press, New York, 17, 371-380.
Isopencu, G. O., Covaliu-Mierlă, C. I., & Deleanu, I. M. (2023). From Plants to Wound Dressing and Transdermal Delivery of Bioactive Compounds. Plants (Basel, Switzerland), 12(14). https://doi.org/10.3390/plants12142661
Kaczmarek-Szczepańska, B., Glajc, P., Chmielniak, D., Gwizdalska, K., Swiontek Brzezinska, M., Dembinska, K., ... & Zasada, L. (2025). Development and Characterization of Biocompatible Chitosan-Aloe Vera Films Functionalized with Gluconolactone and Sorbitol for Advanced Wound Healing Applications. ACS Applied Materials & Interfaces, 17(10). https://doi.org/10.1021/acsami.5c00715
Liang, J., Cui, L., Li, J., Guan, S., Zhang, K., & Li, J. (2021). Aloe vera: a medicinal plant used in skin wound healing. Tissue Engineering Part B: Reviews, 27(5), 455-474. https://doi.org/10.1089/ten.teb.2020.0236
Lowry, O. H., Rosebrough, N. J., Farr, A. L., & Randall, R. J. (1951). Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry, 193(1), 265-275. https://doi.org/10.1016/S0021-9258(19)52451-6
Mahesh, G., Anil Kumar, K., & Reddanna, P. (2021). Overview on the discovery and development of anti-inflammatory drugs: should the focus be on synthesis or degradation of PGE2?. Journal of Inflammation Research, 253-263. https://doi.org/10.2147/JIR.S278514
Mohsin, F., Javaid, S., Tariq, M., & Mustafa, M. (2024). Molecular immunological mechanisms of impaired wound healing in diabetic foot ulcers (DFU), current therapeutic strategies and future directions. International Immunopharmacology, 139. https://doi.org/10.1016/j.intimp.2024.112713
Muller, M. J., Hollyoak, M. A., Moaveni, Z., Brown, T. L. H., Herndon, D. N., & Heggers, J. P. (2003). Retardation of wound healing by silver sulfadiazine is reversed by Aloe vera and nystatin. Burns, 29(8), 834-836. https://doi.org/10.1016/s0305-4179(03)00198-0
Negahdari, S., Galehdari, H., Kesmati, M., Rezaie, A., & Shariati, G. (2017). Wound healing activity of extracts and formulations of aloe vera, henna, adiantum capillus-veneris, and myrrh on mouse dermal fibroblast cells. International Journal of Preventive Medicine, 8(1), 18. https://doi.org/10.4103/ijpvm.IJPVM_338_16
Okur, M. E., Karadağ, A. E., Özhan, Y., Sipahi, H., Ayla, Ş., Daylan, B., ... & Demirci, F. (2021). Anti-inflammatory, analgesic and in vivo-in vitro wound healing potential of the Phlomis rigida Labill. extract. Journal of Ethnopharmacology, 266. https://doi.org/10.1016/j.jep.2020.113408
Patitucci, F., Motta, M. F., Mileti, O., Dattilo, M., Malivindi, R., Pezzi, G., ... & Puoci, F. (2025). Enhancing burn wound care with pre-crosslinked 3D-printed patches: Bromelain delivery and aloe vera bioactives integration for improved healing outcomes. International Journal of Pharmaceutics. https://doi.org/10.1016/j.ijpharm.2025.125304
Rasband, W.S. ImageJ, U. S. (1997). National Institutes of Health, Bethesda, Maryland, USA, 1997-2018. https://imagej.nih.gov/ij/
Re, R., Pellegrini, N., Proteggente, A., Pannala, A., Yang, M., & Rice-Evans, C. (1999). Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biology and Medicine, 26(9-10), 1231-1237. https://doi.org/10.1016/s0891-5849(98)00315-3
Richardson, R., Slanchev, K., Kraus, C., Knyphausen, P., Eming, S., & Hammerschmidt, M. (2013). Adult zebrafish as a model system for cutaneous wound-healing research. Journal of Investigative Dermatology, 133(6), 1655-1665. https://doi.org/10.1038/jid.2013.16
Saeed, M. K., Zahra, N., Abidi, S. H. I., & Syed, Q. U. A. (2022). Phytochemical screening and DPPH free radical scavenging activity of Aloe vera (Aloe barbadensis Miller) powder. International Journal of Food Science and Agriculture, 6(3). http://dx.doi.org/10.26855/ijfsa.2022.09.010
Sena, C. L., Konig, I., da Silva, M. H., Carapiá, M. S., Braga, M. A., & Marcussi, S. (2025). Effects of Uncaria tomentosa and Uncaria guianensis aqueous extract on enzyme activity and modulation of hemostasis. Scientia Plena, 21(2). https://doi.org/10.14808/sci.plena.2025.021101
Singleton, V. L., Orthofer, R., & Lamuela-Raventós, R. M. (1999). [14] Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. In Methods in Enzymology. 299, 152-178.Academic press. https://doi.org/10.1016/S0076-6879(99)99017-1
Smith, J., & Rai, V. (2024). Novel factors regulating proliferation, migration, and differentiation of fibroblasts, keratinocytes, and vascular smooth muscle cells during wound healing. Biomedicines, 12(9). https://doi.org/10.3390/biomedicines12091939
Sumner, J. B., & Graham, V. A. (1921). Dinitrosalicylic acid: a reagent for the estimation of sugar in normal and diabetic urine. Journal of Biological Chemistry, 47(1), 5-9. https://doi.org/10.1016/S0021-9258(18)86093-8
Tomic, D., Shaw, J. E., & Magliano, D. J. (2022). The burden and risks of emerging complications of diabetes mellitus. Nature Reviews Endocrinology, 18(9), 525-539. https://doi.org/10.1038/s41574-022-00690-7
Wang, X., Yang, J., Zhao, Q., Xie, X., Deng, F., Wang, Z., ... & Lv, D. (2024). A tissue-adhesive, mechanically enhanced, natural Aloe Vera-based injectable hydrogel for wound healing: Macrophage mediation and collagen proliferation. International Journal of Biological Macromolecules, 283. https://doi.org/10.1016/j.ijbiomac.2024.137452
Wilkinson, H. N., & Hardman, M. J. (2020). Wound healing: cellular mechanisms and pathological outcomes. Open Biology, 10(9). https://doi.org/10.1098/rsob.200223
Woisky, R. G., & Salatino, A. (1998). Analysis of propolis: some parameters and procedures for chemical quality control. Journal of Apicultural Research, 37(2), 99-105. https://doi.org/10.1080/00218839.1998.11100961
World Health Organization (WHO). (1998). Quality control methods for medicinal plant materials. World Health Organization. Retrieved from: https://www.who.int/docs/default-source/medicines/norms-and-standards/guidelines/quality-control/quality-control-methods-for-medicinal-plant-materials.pdf?sfvrsn=b451e7c6_0. Accessed on 10th August, 2024.
Yousif, D., Yousif, Z., & Joseph, P. (2024). Diabetic Foot Ulcer Neuropathy, Impaired Vasculature, and Immune Responses. In Diabetic Foot Ulcers-Pathogenesis, Innovative Treatments and AI Applications. IntechOpen. https://doi.org/10.5772/intechopen.1003834
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Manhas, D., & Malairaman, U. (2025). Exploring the Potential of Aloe Vera Hydro-Alcoholic Leaf Extract for Topical Diabetic Wound Healing Treatment. International Journal of Advancement in Life Sciences Research, 8(2), 152-168. https://doi.org/https://doi.org/10.31632/ijalsr.2025.v08i02.012
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