Virginia C. A. Martins

Thermal and Rheological Behavior of Collagen. Chitosan blends

Collagen:chitosan blends in 1:1 ratio were prepared and characterized by Fourier transform infrared spectroscopy, thermal (DSC, TG) and rheological studies. Apparently each material maintains its behavior and addition of chitosan does not denature collagen fibers. The rheological behavior showed that adding chitosan to collagen causes a decrease of storage modulus (G’),viscous loss modulus (G”) and apparent viscosity when measured as a function of frequency. Both anionic and native collagen presented more solid-like behavior than fluid-like viscoelastic behavior. Collagen:chitosan blends exhibits a more fluid-like viscoelastic behavior.

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.

Porcine skin as a source of biodegradable matrices: alkaline treatment and glutaraldehyde crosslinking

In this work, the modifications promoted by alkaline hydrolysis and glutaraldehyde (GA) crosslinking on type I collagen found in porcine skin have been studied. Collagen matrices were obtained from the alkaline hydrolysis of porcine skin, with subsequent GA crosslinking in different concentrations and reaction times. The elastin content determination showed that independent of the treatment, elastin was present in the matrices. Results obtained from in vitro trypsin degradation indicated that with the increase of GA concentration and reaction time, the degradation rate decreased. From thermogravimetry and differential scanning calorimetry analysis it can be observed that the collagen in the matrices becomes more resistant to thermal degradation as a consequence of the increasing crosslink degree. Scanning electron microscopy analysis indicated that after the GA crosslinking, collagen fibers become more organized and well-defined. Therefore, the preparations of porcine skin matrices with different degradation rates, which can be used in soft tissue reconstruction, are viable.

Influence of collagen addition on the thermal and morphological properties of chitosan/xanthan hydrogels

This study investigates the collagen influence on thermal and morphological characteristics of chitosan/xanthan hydrogels for potential tissue engineering applications. Anionic collagen was prepared by selective hydrolysis of type I collagen found in bovine tendons. Chitosan was obtained from the partial deacetylation of squid pen β-chitin and xanthan was acquired from Fluka. The hydrogels were obtained in different ratios and were characterized by thermal and morphological analysis. FT-IR suggested only electrostatic interactions between NH3(+) groups of chitosan and COO(-) groups of xanthan and collagen. Thermogravimetric curves showed that hydrogels contain a great amount of water (above 98%) and the presence of collagen does not change this characteristic. Freezing-bound water transition in DSC curves was shifted to higher values due to the increase of water/polymer interaction, mainly when different ratios of chitosan and xanthan were used. SEM images showed sheet-form structures with the presence of collagen promoting an increase in pore size.

Hydroxyapatite and a New Fibrin Sealant Derived from Snake Venom as Scaffold to Treatment of Cranial Defects in Rats

Biomaterials are used as a promising alternative to bone grafts, including bioceramics whose composition resembles that of bone and fibrin sealants due to their hemostatic properties. The objective was to evaluate the repair of cranial defects in 40 rats, grafted with hydroxyapatite and a new fibrin sealant derived from snake venom. The animals were divided into four groups: C (control, no graft); Ha (hydroxyapatite); FS (fibrin sealant), and HaFS (hydroxyapatite and fibrin sealant). The animals were euthanized 2 and 6 weeks after surgery and wound area were submitted to analysis. After 2 weeks, immature bone was formed from the borders of the defect and in groups Ha and HaFS, few hydroxyapatite particles were surrounded by new bone. After 6 weeks, the new bone was mature and surrounded several hydroxyapatite particles, without connective tissue interposition and the volume of new bone was higher in HaFS group. The hydroxyapatite in combination with the new fibrin sealant accelerates bone repair.

Feasibility study of collagen membranes derived from bovine pericardium and intestinal serosa for the repair of cranial defects in ovariectomised rats.

The indication of biomaterials has increased substantially in the regenerative therapy of bone defects. However, in addition to evaluating the physicochemical properties of biomaterials, the quality of the recipient tissue is also essential for the osseointegration of implants, as abnormalities in bone metabolism, such as gonadal hormone deficiency, can influence bone healing. This study evaluated the osteoregenerative capacity of collagen membranes derived from bovine pericardium and intestinal serosa in the repair of cranial defects in ovariectomised rats. Thirty female Wistar rats were submitted to surgical creation of a 5-mm cranial bone defect. The rats were divided into a control group (not ovariectomised) and an ovariectomised group. The non-ovariectomised group was divided into three subgroups: control (G1) in which the defect was not filled with the biomaterial, and two subgroups (G2 and G3) that received the bovine pericardium- and serosa-derived collagen membranes, respectively. The ovariectomised group was divided into the same subgroups (G4, G5, and G6). The animals were sacrificed 8 weeks after surgery. The calvaria were removed for macroscopic and radiographic photodocumentation and processed for histomorphometric analysis of bone healing at the surgical site. Macroscopic, radiological, and microscopic analyses demonstrated the biocompatibility of the implanted collagen membranes, as indicated by the absence of infiltration and signs of inflammation at the surgical site. Histologically, discrete immature bone neoformation projecting from the margins of the defect was observed at the surgical site in ovariectomised groups when compared to the non-ovariectomised groups. The volume of newly formed bone was significantly higher in the non-ovariectomised groups (G1: 7.83%±1.32; G2: 21.33%±1.96; and G3: 22.83%±0.98) compared to the respective ovariectomised subgroups (G4: 3.16%±0.75; G5: 16.83%±0.98; and G6: 16.16%±0.75), thus demonstrating the deleterious effects of ovariectomy on bone homeostasis. Higher volumes of newly formed bone were observed in the groups receiving the membrane grafts (G2, G3, G5, and G6) compared to the control groups (G1 and G4). In conclusion, the bilateral ovariectomy compromises the ability to repair bone lesions grafted with osteoconductive biomaterials as in the case of collagen membranes derived from both bovine pericardium and intestinal serosa.

Effect of stem cells seeded onto biomaterial on the progression of experimental chronic kidney disease

Different routes for the administration of bone marrow-derived cells (BMDC) have been proposed to treat the progression of chronic renal failure (CRF). We investigated whether (1) the use of bovine pericardium (BP) as a scaffold for cell therapy would retard the progression of CRF and (2) the efficacy of cell therapy differently impacts distinct degrees of CRF. We used 2/3 and 5/6 models of renal mass reduction to simulate different stages of chronicity. Treatments consisted of BP seeded with either mesenchymal or mononuclear cells implanted in the parenchyma of remnant kidney. Renal function and proteinuria were measured at days 45 and 90 after cell implantation. BMDC treatment reduced glomerulosclerosis, interstitial fibrosis and lymphocytic infiltration. Immunohistochemistry showed decreased macrophage accumulation, proliferative activity and the expression of fibronectin and α-smooth muscle-actin. Our results demonstrate: (1) biomaterial combined with BMDC did retard the progression of experimental CRF; (2) cellular therapy stabilized serum creatinine (sCr), improved creatinine clearance and 1/sCr slope when administered during the less severe stages of CRF; (3) treatment with combined therapy decreased glomerulosclerosis, fibrosis and the expression of fibrogenic molecules; and (4) biomaterials seeded with BMDC can be an alternative route of cellular therapy.

Effect of Human Adipose Tissue Mesenchymal Stem Cells on the Regeneration of Ovine Articular Cartilage

Cell therapy is a promising approach to improve cartilage healing. Adipose tissue is an abundant and readily accessible cell source. Previous studies have demonstrated good cartilage repair results with adipose tissue mesenchymal stem cells in small animal experiments. This study aimed to examine these cells in a large animal model. Thirty knees of adult sheep were randomly allocated to three treatment groups: CELLS (scaffold seeded with human adipose tissue mesenchymal stem cells), SCAFFOLD (scaffold without cells), or EMPTY (untreated lesions). A partial thickness defect was created in the medial femoral condyle. After six months, the knees were examined according to an adaptation of the International Cartilage Repair Society (ICRS 1) score, in addition to a new Partial Thickness Model scale and the ICRS macroscopic score. All of the animals completed the follow-up period. The CELLS group presented with the highest ICRS 1 score (8.3 ± 3.1), followed by the SCAFFOLD group (5.6 ± 2.2) and the EMPTY group (5.2 ± 2.4) (p = 0.033). Other scores were not significantly different. These results suggest that human adipose tissue mesenchymal stem cells promoted satisfactory cartilage repair in the ovine model.

Agregação de cadeias de acetatos de celulose em LiCl/DMAc: avaliação via viscosimetria

Celulose de linter foi acetilada, visando obter acetatos de celulose com diferentes Graus de Substituicao (GS) em meio homogeneo, usando cloreto de litio/N,N-dimetilacetamida (LiCl/DMAc) como sistema de solvente, e anidrido acetico como reagente acetilante. A agregacao entre cadeias de celulose ou acetatos de celulose (GS 0,8, 1,5 e 2,0) em solucao foi avaliada atraves de medidas viscosimetricas. Os resultados mostraram que a formacao de agregados no sistema de solvente utilizado (LiCl/DMAc) e diferente para celulose e acetatos, e dependente da temperatura e do GS, no caso dos acetatos. Este trabalho corresponde a primeira etapa de um estudo em que se pretende preparar filmes de acetatos, assim como de acetatos reforcados com celulose, diretamente a partir de solucoes destes em LiCl/DMAc. Os resultados apresentados permitem uma escolha melhor embasada do intervalo de concentracao mais adequado para preparacao de filmes, a partir de solucoes de acetatos e celulose nesse sistema de solvente.

Determinação da energia de ativação em hidrogéis poliméricos a partir de dados termogravimétricos

Polyelectrolyte hydrogels formed by chitosan/xanthan (QX) and chitosan/xanthan/collagen (QXC) were prepared and thermogravimetric curves at different heating rates were obtained, with the aim of determining kinetic parameters using the Flynn-Wall method. The calculated activation energy was 3.44 kJ.mol-1 (QX ) and 14.84 kJ.mol-1 (QXC), suggesting stronger interactions in QXC hydrogel structure than in the QX hydrogel, probably due to the presence of carboxyl groups of collagen molecules.