Biochemical similarity between cultured chondrocytes and in situ chondrocytes by chemometric analysis from FTIR microspectroscopy

Monica Maribel Mata-Miranda; Adriana Martinez-Cuazitl; Carla Ivonne Guerrero-Robles; Jesus Emmanuel Noriega-Gonzalez; Juan Salvador Garcia-Hernandez; Gustavo Jesus Vazquez-Zapien

doi:10.1016/j.btre.2019.e00391

Biochemical similarity between cultured chondrocytes and in situ chondrocytes by chemometric analysis from FTIR microspectroscopy

Biotechnol Rep (Amst). 2019 Nov 9:24:e00391. doi: 10.1016/j.btre.2019.e00391. eCollection 2019 Dec.

Authors

Monica Maribel Mata-Miranda¹, Adriana Martinez-Cuazitl^{1

2}, Carla Ivonne Guerrero-Robles¹, Jesus Emmanuel Noriega-Gonzalez¹, Juan Salvador Garcia-Hernandez², Gustavo Jesus Vazquez-Zapien¹

Affiliations

¹ Escuela Militar de Medicina, Centro Militar de Ciencias de la Salud, Secretaría de la Defensa Nacional, Ciudad de México, 11200, Mexico.
² Hospital Central Militar, Secretaría de la Defensa Nacional, Ciudad de México, 11200, Mexico.

Abstract

Background aims: Fourier Transform Infrared Micro-spectroscopy (FTIRM) is an emerging tool that obtains images with biochemical information of samples that are too small to be chemically analyzed by conventional Fourier transform infrared (FTIR) spectroscopy techniques. So, the central objective of this project was to study the biochemical similarity between articular and cultured chondrocytes by chemometric analysis from FTIRM.

Methods: Nine samples of knee articular cartilage were obtained; each sample was divided into two fragments, one portion was used for FTIRM characterization in situ, and from another part, chondrocytes were obtained to be cultured (in vitro), which were subjected to an FTIRM to characterize their biomolecular components. The FTIRM spectra were normalized, and the second derivative was calculated. From these data, principal component analysis (PCA) and a chemometric comparison between in situ and cultured chondrocytes were carried out. Finally, the biochemical mapping was conducted obtaining micro-FTIR imaging.

Results: FTIRM spectra of in situ and in vitro chondrocytes were obtained, and different biomolecules were detected, highlighting lipids, proteins, glycosaminoglycans, collagen, and aggrecan. Despite slight differences in the FTIR spectra, the PCA proved the organic similarity between in situ chondrocytes and cultured chondrocytes, which was also observed in the analysis of the ratios related to the degradation of the articular cartilage and collagen. In the same way, the ability of the FTIRM to characterize the molecular biodistribution was demonstrated.

Conclusion: The biochemical composition and biodistribution analysis using FTIRM have been useful for comparing cultured chondrocytes and in situ chondrocytes.

Keywords: ACI, autologous chondrocyte implantation; Biochemical mapping; Biomolecules; Chemometric analysis; Cultured chondrocytes; ECM, extracellular matrix; FTIR Micro-spectroscopy; FTIR, Fourier Transform Infrared; FTIRI, Micro-FTIR images; FTIRM, Fourier Transform Infrared Micro-spectroscopy; GAGs, glycosaminoglycans; MCT, Mercury-Cadmium-Tellurium; OA, osteoarthritis; PCA, principal component analysis; PGs, proteoglycans; SNV, standard normal variate.