Mahmoud Mudalal1,2,3, Zhanqi Wang1,2, Shockry Mustafa3, Yiping Liu1,2,4, Yao Wang1,2,4, Jize Yu1,2,4, Shengnan Wang4,5, Xiaolin Sun6,7,8, Yanmin Zhou9,10,11. 1. Department of Oral Implantology, Hospital of Stomatology, Jilin University, Changchun, 130021, People's Republic of China. 2. Provincial Key Laboratory of Dental Development, Jaw Remodeling and Regeneration, Jilin University, Changchun, 130021, People's Republic of China. 3. Department of Oral and Maxillofacial Surgery and Periodontology, Faculty of Dentistry, The Arab American University, Jenin, 240, Palestine. 4. Department of Dental Implantology, School and Hospital of Stomatology, Jilin University, No. 1500 Qinghua Rd, Chaoyang district, Changchun City, 130021, Jilin Province, People's Republic of China. 5. College of Clinical Medicine, Jilin University, Changchun, 130021, People's Republic of China. 6. Department of Oral Implantology, Hospital of Stomatology, Jilin University, Changchun, 130021, People's Republic of China. sxl2673366@126.com. 7. Provincial Key Laboratory of Dental Development, Jaw Remodeling and Regeneration, Jilin University, Changchun, 130021, People's Republic of China. sxl2673366@126.com. 8. Department of Dental Implantology, School and Hospital of Stomatology, Jilin University, No. 1500 Qinghua Rd, Chaoyang district, Changchun City, 130021, Jilin Province, People's Republic of China. sxl2673366@126.com. 9. Department of Oral Implantology, Hospital of Stomatology, Jilin University, Changchun, 130021, People's Republic of China. zhouym62@126.com. 10. Provincial Key Laboratory of Dental Development, Jaw Remodeling and Regeneration, Jilin University, Changchun, 130021, People's Republic of China. zhouym62@126.com. 11. Department of Dental Implantology, School and Hospital of Stomatology, Jilin University, No. 1500 Qinghua Rd, Chaoyang district, Changchun City, 130021, Jilin Province, People's Republic of China. zhouym62@126.com.
Abstract
BACKGROUND: An in vitro study on rapid culturing method of human gingival fibroblast cells (HGFCs) was established to investigate the potential use of the leukocyte-platelet rich fibrin (L-PRF) in tissue engineering technology, different medical fields, including periodontology and implantology. METHODS: Eight biopsies were obtained from eight different donors and a modified culturing technique was developed to obtain HGFCs. The modified 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide MTT assay was used to compare the cell viability when the modified culturing method was used in comparison to the standard method. Blood samples were collected from the same patients and L-PRF was isolated using a standard protocol. The releases of platelet-derived growth factor-AA and transforming growth factor-beta1 at various time intervals were observed using enzyme-linked immunosorbent assay (ELISA) kit. The proliferative effect of L-PRF on HGFCs was assessed by the cell counting kit-8 assay. RESULTS: A simple and rapid modified method for in vitro HGFC culture yielded a cellular monolayer within three to nine days after cell culture. L-PRF with three-dimensional polymer fibers released growth factors that peaked during the first three hours and continued to produce up to 10 days. The L-PRF presented a dose-dependent effect on HGFCs proliferation where HGFCs proliferation increased with an increase in L-PRF concentration. CONCLUSION: The modified technique for the culture of HGFCs might be useful for the development of future experimental and clinical studies, besides L-PRF has great therapeutic potential in oral surgery fields.
BACKGROUND: An in vitro study on rapid culturing method of human gingival fibroblast cells (HGFCs) was established to investigate the potential use of the leukocyte-platelet rich fibrin (L-PRF) in tissue engineering technology, different medical fields, including periodontology and implantology. METHODS: Eight biopsies were obtained from eight different donors and a modified culturing technique was developed to obtain HGFCs. The modified 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide MTT assay was used to compare the cell viability when the modified culturing method was used in comparison to the standard method. Blood samples were collected from the same patients and L-PRF was isolated using a standard protocol. The releases of platelet-derived growth factor-AA and transforming growth factor-beta1 at various time intervals were observed using enzyme-linked immunosorbent assay (ELISA) kit. The proliferative effect of L-PRF on HGFCs was assessed by the cell counting kit-8 assay. RESULTS: A simple and rapid modified method for in vitro HGFC culture yielded a cellular monolayer within three to nine days after cell culture. L-PRF with three-dimensional polymer fibers released growth factors that peaked during the first three hours and continued to produce up to 10 days. The L-PRF presented a dose-dependent effect on HGFCs proliferation where HGFCs proliferation increased with an increase in L-PRF concentration. CONCLUSION: The modified technique for the culture of HGFCs might be useful for the development of future experimental and clinical studies, besides L-PRF has great therapeutic potential in oral surgery fields.