Glutaraldehyde ga

Agarose (AGA) was purchased from Bio Plus Fine Research Chemicals. Pluronic Acid (F127), Benzoyl Peroxide (BPO), and Ethanol were purchased from Sigma Aldrich. Eschenstr., Taufkirchen, Germany, Methacrylic Acid (MAA) was purchased from DAE JUNG Chemical and Metals Co. Ltd, Gyeonggi-do, Korea.; Glutaraldehyde (GA) was purchased from Merck, Frankfurter Strasse, Darmstadt, Germany. The Cyclophosphamide drug was obtained from Suzho Unit Phrmaceutical, Suzho, China. Distilled water was prepared freshly in Research Lab, The University of Lahore, Lahore, Pakistan.

Polyether sulfone (PES) flat sheet membranes were prepared by non-solvent induced phase separation (NIPS). Chemicals were purchased from Sigma Aldrich (St. Louis, MO, USA): aluminum oxide (Brockmann activity I, Fluka), 2,2′-azobis(2-methylpropionamidine) dihydrochloride (AIBA), lauryldimethylammonia acetate, latex beads (~1 µm, amine-modified, fluorescent red, L2778), latex beads (~1 µm, carboxylate-modified, fluorescent yellow-green, L4655), lysine, 1-methyl-2-pyrrolidone (NMP), potassium persulfate (KPS), styrene. The 2-Aminoethyl methacrylate hydrochloride (AEMA, Acros Organics), polyethylene glycol (PEG, 400 g mol⁻¹, Acros Organics), and sodium bicarbonate were purchased from Thermo Fisher Scientific (Geel, Belgium). Other purchased chemicals: glutaraldehyde (GA, Merck, Darmstadt, Germany), n-hexane (Merck, Darmstadt, Germany), hydrochloric acid solution (0.1 M, VWR, Radnor, DE, USA), polyether sulfone (PES, Ultrason E2010, BASF, Ludwigshafen, Germany), sodium carbonate (anhydrate, VWR, Radnor, DE, USA), sodium hydroxide solution (0.1 M, VWR, Radnor, DE, USA).
All chemicals were used as received. Ultrapure water was taken from a MilliQ-System (Merck Millipore, Billerica, MA, USA). The dialysis membranes used for the bead purification were obtained from Carl Roth (cellulose acetate, Nadir, molecular weight cut-off (MWCO): 10–20 kDa, Wiesbaden, Germany).

All laboratory materials used in this project, including chitosan [M_w = 1,250,000 (avg.), HS code: 3913900099, 75–85% degree of deacetylation], polylactic acid (medium molecular weight), polyurethane (medium molecular weight), polyethylene glycol (low molecular weight, 400), Tween 80 (26 kDa) were purchased from Sigma Chemical Co. Meanwhile, MnCl₂.4H₂O (98%), FeCl₃.6H₂O (98%), NaOH (99%), and glutaraldehyde (GA) were bought from Merck Co. Lastly, chloroform (99%) and formic acid (95%) solvents were sourced from Sigma-Aldrich, Germany.

Type II collagen gel was obtained from bovine cartilage by alkaline treatments. Each batch of cartilage was supplied from the same cattle farm and type II collagen gels with the same characteristics were extracted by following a patented protocol in the Collagen Department, Skin, and Footwear Research Institute [61 ]. Gentamicin was purchased from Fluka (Milwaukee, WI, USA). Type I collagenase extracted from Clostridium histolyticum was procured from Sigma-Aldrich, Darmstadt, Germany, and glutaraldehyde (GA) from Merck, Darmstadt, Germany. The nanoclays were received from Southern Clay Products (Gonzales, TX, USA,).

For the preparation of graphene oxide (GO) according to the modified Hummer’s method, the chemicals including, sulfuric acid (> 98%), ortho- Phosphoric acid (85%), potassium permanganate (KMnO₄), hydrogen peroxide (30%), and hydrochloric acid (37%), were purchased from Merck. Pristine graphite powder was supplied from Fluka Company. The precursors for the synthesis of the functionalized MGO were iron (II) sulfate tetrahydrate (FeSO₄·4H₂O, 99.7%), iron (III) chloride hexahydrate (FeCl₃·6H₂O, 99.0%), ammonia (NH₄OH, 25 wt%), absolute ethanol, 3-aminopropyl triethoxysilane (AP), and glutaraldehyde (GA) and were purchased from Merck. Rhizopus oryzae lipase (light brown powder, ≥ 30,000 U/g) was purchased from Sigma–Aldrich. Reagents for the nano-biocatalysts assay, p-nitrophenyl palmitate (p-NPP), isopropyl alcohol (99.7%), bovine serum albumin (BSA), Coomassie Brilliant Blue G-250, Triton X-100, and Arabic gum were purchased from Merck. BG-11 medium (Blue-Green medium) components, for the cultivation and maintenance of the microalgae Chlorella vulgaris CCAP were obtained from either Merck. All organic solvents and other chemicals purchased from Merck and were of analytical grade.

The BETA scaffold was manufactured, as previously described [21 (link)]. Briefly, the copolymer emulsion of poly(ε-caprolactone) (PCL: Sigma-Aldrich, Saint Louis, MO, USA, Mn 80,000), and gelatin (Type A from porcine skin, Sigma) dissolved in TFE ((2,2,2-trifluoroethanol; Roth) were spin-coated (2000 rpm) and were dried under vacuum (300 mbar). The optimum mixing ratio of 9.35% PCL and 6.34% gelatin [w/v solvent] was used for the co-polymer of PCL/gelatin as previously described [22 (link)]. Membranes were sterilized before cell culture experiments with PBS, ethanol 80%, and UV exposure. Scanning electron microscopy (SEM, Zeiss Crossbeam 340, Carl Zeiss AG, Oberkochen, Germany) was used to study the morphology of the BETA scaffolds. The cells were fixed in 2% v/v glutaraldehyde (GA; Sigma-Aldrich) and then dehydrated in gradient acetone solutions (Sigma-Aldrich) followed by hexamethyldisilazane (HDMS, Sigma-Aldrich) for 15 min. The samples were subsequently sputter-coated with platinum and analyzed at an operating voltage of 2 kV.