The Bacterial cellulose (Nanoskin^®) raw material was provided from Innovatec’s (São Carlos SP, Brazil). Chon- droitin 4-sulfate and hyaluronic acid sodium salt from Streptococcus equi (bacterial glycosaminoglycan polysaccharide) were purchased from Sigma Aldrich. Vegetal stem cells were obtained from Brazillian environment, Waltheria Douradinha.

The material of the plant of interest is collected and induced damage to causing the formation of scar tissue called callus. This tissue consists of totipotent cells, undifferentiated (stem cells) are collected and grown on agar plates to complete differentiation and generation of a homogeneous culture (2 - 10 days).

Cultures of these stem cells are grown in bioreactors and the batch is collected after all the sugar was metabolized. The cells are washed and homogenized to release secondary metabolites. Soluble metabolites in oil and water are collected and, if you need the isomalt-based spraying is performed.

Transmission infrared spectroscopy (FTIR, Perkin Elmer Spectrum 1000)-Influences of hyaluronic acid (HA) and chondroitin 4-sulfate (CS) in bacterial cellulose were analyzed in the range between 250 and 4000 cm⁻¹ and with 2 cm⁻¹ resolution with samples.

Thermogravimetric analysis (TGA) was carried out for biocomposites using a NETZSCH TG 209F1 in oxygen environment, with a heating rate of 10C/min. The temperature range scanned was from 25 celsius degree to 650 celsius degree. The weight of all specimens was maintained around 10 mg.

In vivo analysis-Evaluation-Clinical study at Sharjah Kuwait Hospital under supervision of Dr. Saqer Al Mualla and Dr. Raed Farahat. Evaluation model-Crushed injury with amputation of the index, Middle and ring of the right hand during work with skin loss since 5 days and all wounds shows necrotic tissue with black parts, bad offensive odor and small amount of pus.

Influences of chondroitin 4-sulfate (CS) and hyaluronic acid in bacterial cellulose (BC) were analyzed by ATR- FTIR in the range of 4000 - 2400 cm⁻¹ (Figure 2) with resolution of 2 cm⁻¹. The main features of the bacterial cellulose in mid-infrared spectroscopy are at 3349 cm⁻¹ (sharp band; O-H stretching vibration associated to intramolecular hydrogen bonding), 3240 cm⁻¹ (medium band; OH Stretching characteristic of hydroxyl

bonding in cellulose structure due to Iα crystalline phase), 2896 cm⁻¹ (C-H stretching vibration of alkane and asymmetric CH₂ stretching), 2853 cm⁻¹ (CH₂ symmetric stretching), 1732 cm⁻¹ (stretching vibration of saturated aliphatic aldehyde carbonyl of BC), 1640 cm⁻¹ (bending mode of absorbed water), 1428 cm⁻¹ (CH₂ or O-H in plane bending), 1373 cm⁻¹ (C-H bending), 1340 cm⁻¹ (OH in-plane bending), 1237 cm⁻¹ (O-H bending), 1203 cm⁻¹ (C-O-H in plane bending at C-6), 1163 cm⁻¹(strong band; antisymmetric bridge C-O-C strentching), 1110 cm⁻¹ (strong band; antisymmetric in-phase ring stretching), 1065 cm⁻¹ (strong band; skeletal vibrations involving C-O stretching at C-3), 1035 cm⁻¹ (strong band; skeletal vibrations involving C-O stretching at C-6), 1015 cm⁻¹ (medium band; skeletal vibrations involving C-O stretching), 897 cm⁻¹ (b-glucosidic linkages between the glucose units) and 748 cm⁻¹ (OH stretching characteristic of hydroxyl bonding in cellulose structure due to Iα crystalline phase) [23] - [26] .

It can be observed from Figures 2(a)-(c), the transmittance intensity is different of bacterial cellulose and bacterial cellulose nanocomposites, which means the exposed groups are interacting with bacterial cellulose components. Similar OH stretching (at 2900 cm⁻¹) can be observed in bacterial cellulose/hyaluronic nanocomposites (BC/HA) and chondroitin 4-sulfate nanocomposites (BC/CS), mainly because of the NH₂ interaction with hydroxyl groups (Figures 2(a)-(c)). Besides, changes can be observed in the symmetrical stretching of CH₂ bonds of bacterial cellulose structures at the absorption peak of 1640 cm⁻¹. Another absorption peak was obtained in the range of 1490 cm⁻¹ on both samples, which shows the presence of a carbonyl group in the bacterial cellulose together with bonds corresponding to those of glycoside, including C-O-C at 1162 cm⁻¹ (as in the case of natural cellulose) [23] - [26] . These results clearly show one possible interaction between bacterial cellulose and chondroitin 4-sulfate/hyaluronic acid, mainly by hydrogen interactions between hydroxyl and carbonyl groups.

In order to analyze thermal behavior for bionanocomposites are characterized typical weight loss versus temperature plots. The TG spectrum (Figure 3) shows a weak loss of weight due to the evaporation of water (at temp. 85 celsius) and also quick drop in weight at a temperature of approx. 300 celsius is mainly attributed to thermal depolymerization of cellulose and the cleavage of glycosidic linkages of cellulose [27] [28] , complete degradation of cellulose take place between 275 and 400 celsius [29] [30] .

All system has similar thermal behavior in bacterial cellulose showed significant alterations. A carbonaceous residue was similar in BC membranes, around 0% at 600 celsius, however sample with hyaluronic acid has little differences in thermal behavior than tested others mainly because there is higher hydrogen bond between bacterial cellulose groups (hydroxyl) and hyaluronic acid (acetyl) which changes bacterial cellulose fibers formation and thermal properties (Figure 4).

Patient enter in Hospital on 12/10/2014 under supervision of Dr. Saqer Al Mualla Consultant Head of Plastic Surgeons Dept’ and Dr. Raed Farahat Plastic Surgeon Specialist, the case planed for skin graft after getting good clean granulation tissue. It was performed immediate intervention with antibiotics and clinical protocols for this disease. In 12/15/2014, started treatment with Bacterial cellulose membranes (Nanoskin^®), one membrane of Nanoskin applied to the wound after removal of all air bubbles under dry gauze used as secondary dressing and light pressure bandage over (Figure 5).

After using the Bacterial cellulose material in alternate days, it can be observed a recovery of the edge and bottom of the wound, however, after 1^st week the wound was better and with necrotic, so, debridement was done and applied Nanoskin (Figure 6).

After, in 2^nd week, wound getting better with minimal exudate and better wound bed. The edge looks health and normal health margin, healthy granulation tissue and no odor and decrease the size of wound bed (Figure 7).

The simple application of dressing, only required the association of saline, gauze and bandage, decrease patient stay and operating room use, resulting in a better cost-benefit. All wounds gets better with Nanoskin^® dressing, dramatic changes was noted after application of Nanoskin^® dressing, patient rarely complain of pain. Healing happened by normal health tissue with less fibrous tissue, normal skin and no pigmentation, besides fast epithelization was observed. The wound healed in 19 days, then the wound coming better even Ring of the hand healed completely after 5 weeks (Figure 8).