CC BY-NC-ND 4.0 · Eur J Dent 2018; 12(03): 344-349
DOI: 10.4103/ejd.ejd_149_17
Original Article
Dental Investigation Society

Cytotoxicity of chlorhexidine and neem extract on cultured human gingival fibroblasts through fluorescence-activated cell sorting analysis : An in-vitro study

Umesh Pratap Verma
1   Department of Periodontology, Faculty of Dental Sciences, King George’s Medical University, Lucknow, Uttar Pradesh, India
,
Abhaya Gupta
1   Department of Periodontology, Faculty of Dental Sciences, King George’s Medical University, Lucknow, Uttar Pradesh, India
,
Rakesh Kumar Yadav
2   Department of Conservative and Endodontics, Faculty of Dental Sciences, King George’s Medical University, Lucknow, Uttar Pradesh, India
,
Rini Tiwari
1   Department of Periodontology, Faculty of Dental Sciences, King George’s Medical University, Lucknow, Uttar Pradesh, India
,
Ramesh Sharma
3   Department of Biochemistry, CSIR.CDRI, Lucknow, Uttar Pradesh, India
,
Anil Kumar Balapure
3   Department of Biochemistry, CSIR.CDRI, Lucknow, Uttar Pradesh, India
› Author Affiliations
Further Information

Publication History

Publication Date:
16 September 2019 (online)

ABSTRACT

Objective: To assess the influence of chlorhexidine (CHX), neem vehicle control (NVC), and neem extract (NE) on cultured human gingival fibroblasts (hGFs) using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and fluorescence-activated cell sorting (FACS) analysis. Materials and Methods: Fibroblasts were derived from healthy gingival biopsy specimens harvested aseptically. The effects of CHX, NVC, and NE were evaluated on cultured hGFs through FACS and MTT assay. Results: MTT assay with hGFs indicated altered morphology with maximum cell death at 10% CHX, while NVC and NE showed similar results at a concentration of 75% and above. On FACS analysis, beyond 1%, CHX adversely affected the cell cycle phase distribution whereas NE exerted a detrimental effect only at 100%. Moreover, both with NVC and NE cells were well differentiated in all the three phases of the cell cycle, with distinction getting lost at 50% to finally causing cell death at 100%. Conclusions: CHX beyond 1% concentration exhibited a toxic effect on hGFs at 1, 5, and 15 min time exposure. However, NE did not adversely affect the fibroblasts even up to 50% concentration showing a less toxic effect in comparison with CHX on these cells. The cytoprotective and oral friendly quality of NE emphaisze the superiority of NE over CHX.

 
  • REFERENCES

  • 1 Cline NV, Layman DL. The effects of chlorhexidine on the attachment and growth of cultured human periodontal cells. J Periodontol 1992; 63: 598-602
  • 2 Mandel ID. Chemotherapeutic agents for controlling plaque and gingivitis. J Clin Periodontol 1988; 15: 488-98
  • 3 Vanka A, Tandon S, Rao SR, Udupa N, Ramkumar P. The effect of indigenous neem Azadirachta Indica [correction of (Adirachta indica)] mouth wash on Streptococcus mutans and Lactobacilli growth. Indian J Dent Res 2001; 12: 133-44
  • 4 Tada H, Shiho O, Kuroshima K, Koyama M, Tsukamato K. An improved colorimetric assay for interleukin-2. J of Immunological methods 1986; 93: 157-165
  • 5 Perry RR, Kang Y, Greaves BR. Relationship between tamoxifen-induced transforming growth factor beta 1 expression, cytostasis and apoptosis in human breast cancer cells. Br J Cancer 1995; 72: 1441-6
  • 6 Kim TK. T test as a parametric statistic. Korean J Anesthesiol 2015; 68: 540-6
  • 7 Damour O, Hua SZ, Lasne F, Villain M, Rousselle P, Collombel C, et al. Cytotoxicity evaluation of antiseptics and antibiotics on cultured human fibroblasts and keratinocytes. Burns 1992; 18: 479-85
  • 8 Chang YC, Huang FM, Tai KW, Chou MY. The effect of sodium hypochlorite and chlorhexidine on cultured human periodontal ligament cells. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2001; 92: 446-50
  • 9 Rajabalian S, Mohammadi M, Mozaffari B. Cytotoxicity evaluation of persica mouthwash on cultured human and mouse cell lines in the presence and absence of fetal calf serum. Indian J Dent Res 2009; 20: 169-73