Hydrogel is being broadly studied due to their tremendous properties, such as swelling behavior and biocompatibility. Numerous review articles have discussed hydrogel polymer types, hydrogel synthesis methods, hydrogel properties, and hydrogel applications. Hydrogel can be synthesized by physical and chemical cross-linking methods. One type of the physical cross-linking method is freeze-thaw (F–T), which works based on the crystallization process of the precursor solution to form a physical cross-link. To date, there has been no review paper which discusses the F–T technique specifically and comprehensively. Most of the previous review articles that exposed the hydrogel synthesis method usually mentioned the F–T process as a small part of the physical cross-linking method. This review attempts to discuss the F–T hydrogel specifically and comprehensively. In more detail, this review covers the basic principles of hydrogel formation in an F–T way, the parameters that influence hydrogel formation, the properties of the hydrogel, and its application in the biomedical field.

A polyvinyl alcohol (PVA) hydrogel loaded with guava leaf extract (GLE) has potential applications as a wound dressing with good antibacterial activity. This study succeeded in fabricating a PVA hydrogel containing GLE using the freeze–thaw (FT) method. By varying the GLE concentration, we can adjust the physical properties of the hydrogel. The addition of GLE results in a decrease in cross-linking during gelation and an increase in the pore size of the hydrogels. The increase of the pore size made the swelling increase and the mechanical strength decrease. The weight loss of the hydrogel also increases because the phosphate buffer saline (PBS) dissolves the GLE. Increasing the GLE concentration caused the Fourier-transform infrared (FTIR) absorbance peaks to widen due to hydrogen bonds formed during the FT process. The crystalline phase was transformed into an amorphous phase in the PVA/GLE hydrogel based on the X-ray diffraction (XRD) spectra. The differential scanning calorimetry (DSC) characterization showed a significant decrease in the hydrogel weight over temperatures of 30–150 °C due to the evaporation of water from the hydrogel matrix. The zone of inhibition of the PVA/GLE hydrogel increased with antibacterial activity against Staphylococcus aureus of 17.93% per gram and 15.79% per gram against Pseudomonas aeruginosa.

The antibacterial activity of garlic (Allium sativum) is believed to be due to its organosulfur compounds, which can supposedly be used further in biomedical applications. This paper reported the use of electrospinning to encapsulate a garlic extract and glycerine in nanofibrous mats. Polyvinylpyrrolidone (PVP) and cellulose acetate (CA) were the building blocks of the composite fibres that served as the hydrophilic matrix to encapsulate the garlic extract with glycerine added mainly to improve the mechanical characteristics of the composite fibres. The combinations of the fibres were PVP/CA, PVP/CA/garlic, PVP/CA/glycerine, and PVP/CA/glycerine/garlic. The characterizations included the morphology, chemical interaction, swelling degree, weight loss, acidity level, wettability, in vitro antibacterial test, and release behaviour test. The composite nanofibrous mats were uniform, bead-free with a size ranging from 350 nm to 900 nm. The Fourier-transform infrared spectra proved the presence of the garlic extract and glycerine in the fibres. The swelling degree test showed that the fibrous mats generally did have maximum swelling degrees above 100% except for the PVP/CA fibrous mat, whose maximum value was not achieved within 48 hours. The fibrous mat with glycerine showed generally larger weight loss compared to the fibrous mats without glycerine. The result of the contact angle measurement proved that the composite fibres are all hydrophilic with the PVP/CA/glycerine/garlic fibres being the least hydrophilic. The pH level of the fibre mats was from 3.7 to 4.0 due to the use of acetic acid. The Young's modulus and ultimate tensile strength of the mats were significantly reduced due to the presence of glycerine. The encapsulation of the garlic extract in the fibres did not eliminate the antibacterial activity of the garlic extract, as proven in the in vitro antibacterial test. The release of the garlic extract from the composite PVP/CA/glycerine/garlic fibres was found to be the largest due to the large diameter of the fibres, while the blend of PVP with CA successfully reduced the rate of release due to the insolubility of CA. We successfully encapsulated the garlic extract and glycerine in the PVP/CA nanofibrous mats with antibacterial activity.

According to the Indonesian Ministry of Health Research and Development Agency's 2018 Basic Health Research (Riskesdas), bone defects, because of fracture cases in Indonesia, reached a prevalence of 5.5%. Bone tissue engineering is one of the most promising new approaches for accelerating the growth and healing of bone defects in patients. In this study, a bone scaffold electrospun nanofiber composed of hydroxyapatite (HA)-polycaprolactone (PCL)-collagen was fabricated to mimic the extracellular matrix (ECM) found in native bone tissue and to promote bone remodeling and healing. Through characterization of functional groups with Fourier transform infrared (FTIR), morphology scanning electron microscopy (SEM), tensile strength test, degradation test, and cytotoxicity MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, this study determines the optimal composition of electrospun nanofiber HA/PCL/collagen to obtain the best candidate of a bone scaffold with excellent characteristics. Electrospinning was used to fabricate bone scaffolds. The samples used in this study consisted of control samples, namely PCL-HA and PCL-Collagen, and three samples with a mass ratio composition of HA-PCL-collagen 50:40:10, 50:30:20, and 50:25:25 (w/v%). The FTIR analysis of the sample revealed that no chemical bond existed between the materials. The HA/PCL/collagen 50:25:25 (F3) sample exhibited the best characteristics as a bone scaffold candidate, with a fiber diameter of 365 ± 202 nm; a porous fraction area of 58.98%; an ultimate tensile strength, and an elasticity modulus of 0.60 ± 0.185 and 5.98 ± 0.82 MPa, respectively; a degradation rate of 1.93 × 10⁻⁶ g/h; and cell viability of 81.03%. Graphical Abstract

Porosity is one of the parameters of scaffold pore structure that must be developed using paraffin wax as a synthetic polymer for making porous bioceramics carbonated hydroxyapatite (CHA). This study fabricated CHA based on abalone mussel shells (Halioitis asinina); CHA/paraffin wax nanocomposite scaffolds were synthesized using paraffin wax with concentration variations of 10, 20, and 30 $\mathrm{wt}.\%$. The energy-dispersive X-ray spectroscopy (EDS) results showed that the Ca/P molar ratio of CHA was 1.72, which approaches the natural bone. The addition of paraffin wax in all concentration variation treatments caused the crystallographic properties of the CHA/paraffin wax nanocomposite scaffolds to decrease. The results of pore analysis suggest that the high concentration of paraffin wax in the CHA suspension is involved in the formation of more pores on the surface of the scaffold, but only CHA/paraffin wax 30 $\mathrm{wt}$.% had a scaffold with potential to be used in media with a cellular growth orientation. The micropore analysis was also supported by the cell viability assay results for CHA/paraffin wax 30 $\mathrm{wt}$.% nanocomposite scaffold, where serial doses of scaffold concentrations to mouse osteoblast cells were secure. Overall, based on this analysis, the CHA/paraffin wax scaffold can be a candidate for bone tissue engineering.