**2.3 Tissue samples**

Fresh cadaver buffalo aorta (**Figure 1A**), buffalo diaphragm (**Figure 1B**) and goat skin (**Figure 1C**) collected in chilled (4°C) sterile 1X PBS (pH 7.4) containing 0.016% gentamicin (antibiotic), 0.0205% EDTA (proteolytic inhibitor) and 0.1% NaN3 (antimycotic) were our study materials. A cortical bone collected from the anterior diaphysis of the right femur of an adult cadaver Gir cow was also used.

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**Figure 2.**

*Fourier Transform Infrared Spectroscopy of the Animal Tissues*

**3.1 Fourier transform infrared spectroscopy of the buffalo aorta**

**3.2 Fourier transform infrared spectroscopy of the buffalo diaphragm**

The diaphragm is a dome shaped structure, composed of muscle surrounding a central tendon, which separates the thoracic and abdominal cavities. Before

*FTIR spectra showing transmittance peaks of native aorta (NA) at 1282.55, 1526.71, 1658.84, 2958.9 and 3282.95 cm−1; and decellularized aorta (DA) at 1230.69, 1529.60, 1658.84, 2954.08 and 3280 cm−1.*

The *aorta,* an elastic artery, has a trilaminar structure consisting of a tunica intima, media, and adventitia. The media comprises cellular elements (including smooth muscle cells) and structural proteins (notably collagen and elastin) that form the ECM. Before being used in regenerative therapies, cellular elements of the aorta should be removed by decellularization process. The decellularization of fresh posterior aorta from deceased donor buffalo was completely achieved by treatment with 1% SDS for 24 hours followed by 0.25% trypsin for 2 hours and again by 1% SDS for 24 hours [19]. Both the native and decellularized aortae were characterized by FTIR spectroscopy [14]. Herein, one milligram of each freeze-dried tissues were mixed with pure dry KBr powder in 1:10 ratio, and pelleted. The FTIR spectra were recorded by an infrared spectrophotometer in the 500–4000 cm−1 wave number spectral range with a spectral resolution of 2 cm−1 and 45 scans. **Figure 2** illustrates the FTIR spectra of native and decellularized aortae. The transmittance peaks indicated the presence of organic collagen amide A, amide B, amide I, amide II and amide III chemical functional groups in both native and decellularized aortae. The amide A band (3294 cm−1) is associated with H-bonded N-H stretching [11, 13, 20] and was found at 3282.95 cm−1 for native aorta and 3280 cm−1 for decellularized aorta. The amide B band (2953 and 2928 cm−1) is related to CH2 asymmetric stretching [11, 13, 21] and was observed at 2958.9 cm−1 for native aorta and 2954.08 cm−1 for decellularized aorta. The amide I band (1641–1658 cm−1) is associated with C=O hydrogen bonded stretching [11, 13, 22] as recorded at 1658.84 cm−1 for native aorta and 1658.84 cm−1 for decellularized aorta. The amide II (1539–1546 cm−1) is associated with C-N stretching and N-H in plane bending from amide linkages, including wagging vibrations of CH2 groups from the glycine backbone and proline sidechains [11, 13, 23] in native aorta and decellularized aorta appeared at 1526.71 cm−1 and 1529.60 cm−1, respectively. The amide III (NH bend) band was found at 1282.55 cm−1 for NA and 1230.69 cm−1 for decellularized aorta [11, 13, 24].

*DOI: http://dx.doi.org/10.5772/intechopen.94582*

**3. Methods**

**Figure 1.** *Gross images of buffalo aorta (a), buffalo diaphragm (B), and goat skin (C).*
