Human gingival fibroblasts culture in an autologous scaffold and assessing its effect on augmentation of attached gingiva in a pilot clinical trial

BACKGROUND AND AIM: An important goal of periodontal plastic surgery is the creation of attached gingiva around the teeth. In this study, the aims were to culture gingival fibroblasts in a biodegradable scaffold and measure the width of attached gingiva after the clinical procedure. METHODS: This study was carried out on 4 patients (8 sites), with inadequate attached gingiva next to at least two teeth in contralateral quadrants of the same jaw. A biopsy of attached gingiva (epithelial + connective tissue) was taken using a surgical blade. Following culture of gingival fibroblasts, 250 × 103 cells in 250 μl nutritional medium were mixed with platelet-rich in growth factor (PRGF). Periosteal fenestration technique was done on one side (control) and tissue-engineered mucosal graft (test) was carried out on the contralateral side in each patient. The width of keratinized tissue, probing depth (PD) and width of attached gingiva were recorded at baseline and 3 months after the operation. RESULTS: An increased width of keratinized and attached tissue on all operated sites after 3 months was observed. These results showed the increased mean of the width of keratinized and attached gingiva to be 4.17 mm and 4.14 mm in test and 1.10 mm and 1.10 mm in control sites, respectively. The difference of keratinized and attached gingiva width between test and control sites was significant (P = 0.030, and P = 0.010 respectively). CONCLUSION: According to the results of this study, PRGF can be used as a scaffold to transfer gingival fibroblasts to recipient sites with significant clinical results. KEYWORDS: Tissue Engineering; Gingiva; Blood Platelet; Scaffol

Cytotoxicity evaluation of Persica mouthwash on cultured human and mouse cell lines in the presence and absence of fetal calf serum

Background and Aims: The effectiveness of an ideal antimicrobial agent depends on its ability to kill microbes while causing minimal toxicity to host cells. Several studies have been reported on the antimicrobial effects of chewing sticks (Salvadora persica) on oral bacteria. The purpose of this study was to evaluate the cytotoxic effects of Persica™ and chlorhexidine (CHX) mouthwashes on cultured human and mouse cell lines. Materials and Methods: This was an experimental study. The toxic effects of four dilutions of Persica™ and CHX mouthwashes on KB, Saos-2, J744 A1, and gingival fibroblast cells were evaluated by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay. The effect of fetal calf serum (FCS) components on the cytotoxicity of these mouthwashes was also investigated. Statistical Analysis: Analysis of variance and the Kruskal-Wallis test were used to evaluate the results. Results: The results indicated that Persica™, at concentrations higher than 0.1%, exerted a very significant cytotoxic effect on all the cell lines (P ≤ 0.01). CHX, at a concentration of 0.001%, exerted toxic effects only on gingival fibroblasts; concentrations higher than 0.001% were required to produce significant cell death in the other cell lines. At all the concentrations under study, both Persica™ and CHX exerted significantly greater cytotoxic effects in the absence of FCS than in its presence (i.e., in control culture medium). The toxicities of both mouthwashes were attenuated in the presence of FCS (10%). Conclusion: Our results indicate that both Persica™ and CHX mouthwashes are toxic to macrophage, epithelial, fibroblast, and osteoblast cells in a concentration-dependent manner

Evaluation of effects of diclofenac on the proliferation and differentiation of PC12 cells in vitro

Background & Objective: Diclofenac is a non-steroidal, anti-inflamatory drug that is prescribed as an analgesic. However, there is little known about the effects of diclofenac on the neural cells. In this study, we investigated the effects of diclofenac as sodium salt on the proliferation and differentiation of PC12 cells.   Materials & Methods: This expeimental study was done in Kerman neuroscience research center during 2004. The cell proliferation was evaluated by using XTT assay in the both free-serum neurobasal medium supplemented with B27 supplement and DMEM/F12 medium containing 10% FBS. The nerve growth factor(NGF) – induced differentiation was assessed  by measuring the neurite length for each treatment.   Results: The drug toxicity was exhibited at the higher concentrations of 310 mM in the supplemented neurobasal medium. The treatment of cells in the DMEM/F12 medium increased their sensitivity to diclofenac, with 40 and 85% growth inhibition at the 155 and 310 mM concentrations, respectively. The different generics of drug exhibited a equal toxic effects on the PC12 cells. The NGF- induced differentiation was not reduced by toxic and subtoxic concentrations of diclofenac.   Conclusion: This study indicated that diclofenac may be able to exhibit its neurotoxic effects through growth inhibition, but not differentiation inhibition. B27 supplement has several antioxidant compounds. Therefore, the difference of diclofenac cytotoxic effects in two culture media suggest that drug cytotoxicity may be related to the oxidative stress

Human gingival fibroblasts culture in an autologous scaffold and assessing its effect on augmentation of attached gingiva in a pilot clinical trial

BACKGROUND AND AIM: An important goal of periodontal plastic surgery is the creation of attached gingiva around the teeth. In this study, the aims were to culture gingival fibroblasts in a biodegradable scaffold and measure the width of attached gingiva after the clinical procedure. METHODS: This study was carried out on 4 patients (8 sites), with inadequate attached gingiva next to at least two teeth in contralateral quadrants of the same jaw. A biopsy of attached gingiva (epithelial + connective tissue) was taken using a surgical blade. Following culture of gingival fibroblasts, 250 × 103 cells in 250 μl nutritional medium were mixed with platelet-rich in growth factor (PRGF). Periosteal fenestration technique was done on one side (control) and tissue-engineered mucosal graft (test) was carried out on the contralateral side in each patient. The width of keratinized tissue, probing depth (PD) and width of attached gingiva were recorded at baseline and 3 months after the operation. RESULTS: An increased width of keratinized and attached tissue on all operated sites after 3 months was observed. These results showed the increased mean of the width of keratinized and attached gingiva to be 4.17 mm and 4.14 mm in test and 1.10 mm and 1.10 mm in control sites, respectively. The difference of keratinized and attached gingiva width between test and control sites was significant (P = 0.030, and P = 0.010 respectively). CONCLUSION: According to the results of this study, PRGF can be used as a scaffold to transfer gingival fibroblasts to recipient sites with significant clinical results