Moghaddas Omid, Miremadi Behdokht, Seyed Jafari Ehsan
Department of Periodontics, Faculty of Dentistry, Tehran Medical Sciences Islamic Azad University, Tehran, Iran.
Private Practice, Tehran, Iran.
J Adv Periodontol Implant Dent. 2020 Jul 5;12(2):84-89. doi: 10.34172/japid.2020.011. eCollection 2020.
Periodontal regeneration is an essential goal of periodontal therapy. Acellular dermal matrix (ADM) has been recommended as an alternative to autogenous grafts. However, since it is devoid of cells and vasculature, there are concerns regarding the biological behavior of cells on ADM. This study aimed to assess the effects of two commonly used ADMs on biological behavior, i.e., attachment and proliferation, of human gingival fibroblasts (HGFs).
This in vitro, experimental study was conducted on explanted and cultured HGFs. ADM types 1 and 2 (n=26; measuring 10×15 mm) were rinsed with saline solution, adapted to the bottom of 52 wells, exposed to HGFs with a cell density of 16,000 cells/mL, and incubated at 37°C for 12, 24, and 84 hours and seven days. Cell attachment was assessed 12 hours after incubation using 4›,6-diamidino-2-phenylindole (DAPI) and methyl-thiazol-diphenyl-tetrazolium (MTT) assay under a fluorescence microscope. Cell viability was assessed at 24 and 84 hours and one week using the MTT assay. Cells were then platinum-coated, and their morphology was evaluated under a scanning electron microscope (SEM). Data were analyzed using ANOVA.
HGFs were evaluated in 60 samples in three groups (n=20). Cell attachment was the same in the three groups, as shown by the MTT assay and DAPI test (P=0.6). Cell viability at one week was 3.73±0.02, 2.88±0.29, and 2.13±0.24 in the control, ADM 1, and ADM 2 groups, respectively. The difference was statistically significant (P=0.01).
Both scaffolds were the same in terms of attachment of HGFs. However, ADM 1 was superior to ADM2 in terms of cell viability and morphology at one week. It was concluded that the quality of acellular dermal scaffolds could significantly influence cellular behaviors and tissue maturation.
牙周组织再生是牙周治疗的重要目标。脱细胞真皮基质(ADM)已被推荐作为自体移植物的替代物。然而,由于其缺乏细胞和血管,人们对细胞在ADM上的生物学行为存在担忧。本研究旨在评估两种常用的ADM对人牙龈成纤维细胞(HGFs)生物学行为(即附着和增殖)的影响。
本体外实验研究对原代培养的HGFs进行。将1型和2型ADM(n = 26;尺寸为10×15 mm)用盐溶液冲洗,放置于52个孔的底部,接种细胞密度为16,000个细胞/mL的HGFs,并在37°C下孵育12、24、84小时和7天。孵育12小时后,使用4',6-二脒基-2-苯基吲哚(DAPI)和甲基噻唑基二苯基四氮唑溴盐(MTT)法在荧光显微镜下评估细胞附着情况。在24、84小时和1周时使用MTT法评估细胞活力。然后对细胞进行铂包被,并在扫描电子显微镜(SEM)下评估其形态。数据采用方差分析进行分析。
在三组(n = 20)的60个样本中对HGFs进行了评估。MTT法和DAPI检测显示,三组中的细胞附着情况相同(P = 0.6)。对照组、ADM 1组和ADM 2组在1周时的细胞活力分别为3.73±0.02、2.88±0.29和2.13±0.24。差异具有统计学意义(P = 0.01)。
两种支架在HGFs附着方面相同。然而,在1周时,ADM 1在细胞活力和形态方面优于ADM 2。得出的结论是,脱细胞真皮支架的质量可显著影响细胞行为和组织成熟。