Local tissue reaction and histopathological characteristics of three different bulking agents: a rabbit model

Abstract

Purpose: We assessed the efficacy and safety of a single injection of three bulking agents over the short- and long-term follow-ups in rabbits. Dermal and preputial matrices were compared with Deflux (DxHA) injection.

Material and methods: Twenty-four rabbits were divided into three groups. Group I (n=8) underwent the injection of a lyophilized dermal matrix (LDM) beneath the seromuscular layer of the bladder wall. Rabbits in group II (n=8) were injected with lyophilized preputial matrix (LPM). Rabbits of group III (n=8) were injected with DxHA as the control group. They were followed up for 1 and 6 months after the injection. Subcutaneous injection of all bulking agents was also performed in nude mice. Biopsies were stained with LCA (leukocyte common antibody), CD68, CD31, and CD34. Scanning electron microscopy (SEM) and MTT assay were also performed.

Results: Immunohistochemistry staining with CD68 and LCA revealed higher inflammation grade in LDM as compared with LPM and DxHA. Fibrosis grade was also higher in LDM both in short- and long-term follow-ups. However, no significant difference was detected in CD31 and CD34 staining between control and experimental groups. SEM analysis showed that the particle size of LPM was more similar to DxHA. MTT assay revealed that cell proliferation was similar in DxHA, LDM, and LPM. In-vivo assay in nude mice model showed more promising results in LPM as compared with LDM.

Conclusion: The long-term results demonstrated that LPM was more similar to Deflux with the least local tissue reaction, inflammation, and fibrosis grade.

Keywords: Acellular Dermis; Bulkamid [Supplementary Concept]; dextranomer [Supplementary Concept].

Figure 1. LPM and LDM supplied in the syringe at the time of injection (A) Injection method of Deflux, LPM, and LDM beneath the mucosa layer of the bladder wall (B).

Figure 2. H&E staining: Decellularized preputial (A) and skin tissues (B).

Figure 3. SEM analysis of LDM (A), LPM (B), and Deflux particles (C) before injection.

Figure 4. IHC staining after 1 month of follow-up: CD31 staining in Deflux (A), LDM (B), and LMP (C), CD34 staining in Deflux (D), LDM (E), and LMP (F), CD68 staining in Deflux (G), LDM (H), and LMP (I), and LCA staining in Deflux (J), LDM (K), and LMP (L). IHC staining after 6 months of follow-up: CD31 staining in Deflux (M), LDM (N), and LMP (O), CD34 staining in Deflux (P), LDM (Q), and LMP (R), CD68 staining in Deflux (S), LDM (T), and LMP (U), and LCA staining in Deflux (V), LDM (W), and LMP (X).