Bacterial Cellulose/Collagen Hydrogel for Wound Healing

This study compares a wound dressing based on bacterial cellulose/collagen (BC/COL) hydrogel in rat dorsum with commercial collagenase ointment and untreated wound. The hydrogel was characterized by Scanning Electron Microscopy (SEM), Thermogravimetric Analysis (TGA), Fourier transformed - Infrared spectroscopy (FT-IR) and X-ray diffraction (XRD). According to the In vivo test and macroscopic evaluation, BC/COL hydrogel showed a better repair of wounds and promoted statistically significant differences of tissue repair between treatments on the 7th day after surgery. Better quality, quantity and orientation evaluation of collagen fibers (p=0.0001) were observed in the BC/COL hydrogel and collagenase ointment groups in relation to the control group. The BC/COL hydrogel promoted better wound healing than collagenase and the control group, therefore, it can be considered a potential wound dressing for skin regeneration.

Keywords:
bacterial cellulose; collagen; hydrogel; wound dressing; tissue repair

Authorship

Paula Rodrigues Fontes de Sousa Moraes ^**E-mail: paularfsm@usp.br

Programa de Pós Graduação Interunidades Bioengenharia- EESC/FMRP/IQSC- USP, Av. Trabalhador São-carlense, 400, Arnold Schimidt, CEP: 13566-590, São Carlos, SP, BrazilUSPBrazilSão Carlos, SP, BrazilPrograma de Pós Graduação Interunidades Bioengenharia- EESC/FMRP/IQSC- USP, Av. Trabalhador São-carlense, 400, Arnold Schimidt, CEP: 13566-590, São Carlos, SP, Brazil

Sybele Saska

Instituto de Química – Universidade Estadual Paulista - UNESP, Rua Prof. Francisco Degni, 55, Quitandinha, CEP: 14800-060, Araraquara, SP, BrazilUniversidade Estadual PaulistaBrazilAraraquara, SP, BrazilInstituto de Química – Universidade Estadual Paulista - UNESP, Rua Prof. Francisco Degni, 55, Quitandinha, CEP: 14800-060, Araraquara, SP, Brazil

Hernane Barud

Laboratório de Biopolímeros e Biomateriais (BioPolMat), Centro Universitário de Araraquara – UNIARA, Rua Voluntários da Pátria, 1309, Centro, CEP: 14801-320, Araraquara, SP, BrazilCentro Universitário de AraraquaraBrazilAraraquara, SP, BrazilLaboratório de Biopolímeros e Biomateriais (BioPolMat), Centro Universitário de Araraquara – UNIARA, Rua Voluntários da Pátria, 1309, Centro, CEP: 14801-320, Araraquara, SP, Brazil

Laís Roncalho de Lima

Virgínia da Conceição Amaro Martins

Instituto de Química de São Carlos/USP, Av. Trabalhador São-carlense, 400, Arnold Schimidt, CEP: 13566-590, São Carlos, SP, BrazilUSPBrazilSão Carlos, SP, BrazilInstituto de Química de São Carlos/USP, Av. Trabalhador São-carlense, 400, Arnold Schimidt, CEP: 13566-590, São Carlos, SP, Brazil

Ana Maria de Guzzi Plepis

Sidney José Lima Ribeiro

Ana Maria Minarelli Gaspar

Laboratório de Morfologia, Universidade Estadual Paulista - UNESP, Rua Humaitá, 1680, Centro, CEP: 14801-903, Araraquara, SP, BrazilUniversidade Estadual PaulistaBrazilAraraquara, SP, BrazilLaboratório de Morfologia, Universidade Estadual Paulista - UNESP, Rua Humaitá, 1680, Centro, CEP: 14801-903, Araraquara, SP, Brazil

*E-mail: paularfsm@usp.br

SCIMAGO INSTITUTIONS RANKINGS

Figures | Tables

Figures (9)
Tables (2)

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Figure 1
SEM surface images of pure collagen (a) and BC/COL hydrogel (c). SEM crosssection images of pure collagen (b) and BC/COL hydrogel (d).

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Figure 2
TG curves (----) and DTG (-----): (a) pure collagen, (b) BC/COL hydrogel and (c) pristine BC.

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Figure 3
XRD diffraction patterns of the BC/COL hydrogel (a), pure collagen (b) and pristine BC (c).

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Figure 4
FT-IR spectra: (a) BC/COL hydrogel, (b) pure collagen and (c) pristine BC.

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Figure 5
Values measured for the tissue repair of wounds of the GI, GII and GIII groups for experimental periods of 3, 7, 15 and 30 days. Values are expressed as mean ± standard error; * p<0.05. The one-way ANOVA followed by the post-hoc t test (LSD) were applied.

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Figure 6
3-day period, (a) – GI shows early neovascularization, crust and disorganized granulation tissue and moderate inflammatory reaction in the epithelial healing process; (b) – GII shows absence of epithelium, crust and disorganized granulation tissue and mild inflammatory reaction; (c) – GIII shows absence of epithelium, disorganized granulation tissue and mild inflammatory reaction. Inflammatory infiltrates were predominantly composed of polymorphonuclear cells, such as macrophages and neutrophils. Epithelium (A), crust (B), neovascularization (C), inflammatory infiltrate (D) and granulation tissue (E). Staining: H&E.

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Figure 7
7-day period, (a) – GI shows early neovascularization, crust, granulation tissue with early organization of collagen fibers and moderate inflammatory reaction in the epithelial healing process; (b) – GII shows absence of epithelium, early neovascularization, crust, moderate inflammatory reaction with early organization of collagen fibers; (c) – GIII shows absence of epithelium, early neovascularization, disorganized granulation tissue and moderate inflammatory reaction. Epithelium (A), crust (B), neovascularization (C), inflammatory infiltrate (D) and organization of collagen fibers (E). Staining: H&E.

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Figure 8
15-day period, (a) – GI shows complete epithelium and dermis repair, neovascularization, keratin layer and mild inflammatory reaction; (b) - GII shows epithelium in formation and dermis repair, keratin layer, neovascularization and mild inflammatory reaction; (c) – GIII shows re-epithelialization process, keratin layer, disorganized granulation tissue and moderate inflammatory reaction. Epithelium (A), keratin layer (B), inflammatory infiltrate (C), neovascularization (D) and organization of collagen fibers (E). Staining: H&E.

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Figure 9
30-day period, (a) – GI shows complete epithelium and dermis repair, keratin layer and hair follicle; (b) – GII shows complete epithelium and dermis repair with hair follicle and glands; (c) – GIII shows organized tissue, re-epithelialization, keratin layer and presence of glands. Epithelium (A), keratin layer (B), neovascularization (C), organization of collagen fibers (D), glands (E) and hair follicle (F). Staining: H&E.

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Table 1
Classification of the tissue repair pattern regarding quality, quantity and orientation of collagen fibers.

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Table 2
Classification of the tissue repair pattern in relation to the epithelial repair.

Figure 1 SEM surface images of pure collagen (a) and BC/COL hydrogel (c). SEM crosssection images of pure collagen (b) and BC/COL hydrogel (d).

Figure 2 TG curves (----) and DTG (-----): (a) pure collagen, (b) BC/COL hydrogel and (c) pristine BC.

Figure 3 XRD diffraction patterns of the BC/COL hydrogel (a), pure collagen (b) and pristine BC (c).

Figure 4 FT-IR spectra: (a) BC/COL hydrogel, (b) pure collagen and (c) pristine BC.

Figure 5 Values measured for the tissue repair of wounds of the GI, GII and GIII groups for experimental periods of 3, 7, 15 and 30 days. Values are expressed as mean ± standard error; * p<0.05. The one-way ANOVA followed by the post-hoc t test (LSD) were applied.

Figure 6 3-day period, (a) – GI shows early neovascularization, crust and disorganized granulation tissue and moderate inflammatory reaction in the epithelial healing process; (b) – GII shows absence of epithelium, crust and disorganized granulation tissue and mild inflammatory reaction; (c) – GIII shows absence of epithelium, disorganized granulation tissue and mild inflammatory reaction. Inflammatory infiltrates were predominantly composed of polymorphonuclear cells, such as macrophages and neutrophils. Epithelium (A), crust (B), neovascularization (C), inflammatory infiltrate (D) and granulation tissue (E). Staining: H&E.

Figure 7 7-day period, (a) – GI shows early neovascularization, crust, granulation tissue with early organization of collagen fibers and moderate inflammatory reaction in the epithelial healing process; (b) – GII shows absence of epithelium, early neovascularization, crust, moderate inflammatory reaction with early organization of collagen fibers; (c) – GIII shows absence of epithelium, early neovascularization, disorganized granulation tissue and moderate inflammatory reaction. Epithelium (A), crust (B), neovascularization (C), inflammatory infiltrate (D) and organization of collagen fibers (E). Staining: H&E.

Figure 8 15-day period, (a) – GI shows complete epithelium and dermis repair, neovascularization, keratin layer and mild inflammatory reaction; (b) - GII shows epithelium in formation and dermis repair, keratin layer, neovascularization and mild inflammatory reaction; (c) – GIII shows re-epithelialization process, keratin layer, disorganized granulation tissue and moderate inflammatory reaction. Epithelium (A), keratin layer (B), inflammatory infiltrate (C), neovascularization (D) and organization of collagen fibers (E). Staining: H&E.

Figure 9 30-day period, (a) – GI shows complete epithelium and dermis repair, keratin layer and hair follicle; (b) – GII shows complete epithelium and dermis repair with hair follicle and glands; (c) – GIII shows organized tissue, re-epithelialization, keratin layer and presence of glands. Epithelium (A), keratin layer (B), neovascularization (C), organization of collagen fibers (D), glands (E) and hair follicle (F). Staining: H&E.

Table 1 Classification of the tissue repair pattern regarding quality, quantity and orientation of collagen fibers.

Classification	Score
Absence	0
Disorganized fibers	1
Poorly organized fibers	2
Medium organization of fibers	3
Well organized fibers	4

Table 2 Classification of the tissue repair pattern in relation to the epithelial repair.

Classification	Score
Unrepaired	0
Healing process	1
Repaired	2

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[1] *E-mail: paularfsm@usp.br