In this study, we developed a composite filler comprising cross-linked hyaluronic acid (HA) and human collagen (COL) derived from the human umbilical cord with the aim of improving its biocompatibility and longevity compared with commercially available fillers. After HA/COL composite fillers were ma...
In this study, we developed a composite filler comprising cross-linked hyaluronic acid (HA) and human collagen (COL) derived from the human umbilical cord with the aim of improving its biocompatibility and longevity compared with commercially available fillers. After HA/COL composite fillers were made in two different ratios (10:1 and 5:1), the physical properties of the fillers were evaluated. The interior morphologies and in vivo weight change of these hydrogels were also characterized at 1-16 weeks after injection into mice. To evaluate their biocompatibility and durability in vivo, we injected the composite fillers into nude mice subcutaneously. The variations of injected gel weight were measured and compared with the commercial dermal fillers (Restylane and TheraFill). The composites showed improved or similar physical properties (complex viscosity of 19-22 × 105 cP, and injection force of 10-12 N) over the commercial dermal fillers. Sixteen weeks following the injection, the ratio of remaining composite filler weight to initial weight (75.5 ± 16.9%; 10:1) was shown to be greater than that of the commercial fillers (43.2 ± 8.1%, Restylane; 12.3 ± 5.3%, TheraFill). In addition, immunohistochemical analysis with angiogenesis-related markers such as isolectin and vWF revealed newly formed blood vessels and cellular influx into the composite filler, which were not observed in the other fillers. These results clearly suggest that the HA/COL composite filler is a superior candidate for soft tissue reconstruction. The filler we developed may be a suitable candidate as an injectable dermal filler for tissue augmentation in humans.
In this study, we developed a composite filler comprising cross-linked hyaluronic acid (HA) and human collagen (COL) derived from the human umbilical cord with the aim of improving its biocompatibility and longevity compared with commercially available fillers. After HA/COL composite fillers were made in two different ratios (10:1 and 5:1), the physical properties of the fillers were evaluated. The interior morphologies and in vivo weight change of these hydrogels were also characterized at 1-16 weeks after injection into mice. To evaluate their biocompatibility and durability in vivo, we injected the composite fillers into nude mice subcutaneously. The variations of injected gel weight were measured and compared with the commercial dermal fillers (Restylane and TheraFill). The composites showed improved or similar physical properties (complex viscosity of 19-22 × 105 cP, and injection force of 10-12 N) over the commercial dermal fillers. Sixteen weeks following the injection, the ratio of remaining composite filler weight to initial weight (75.5 ± 16.9%; 10:1) was shown to be greater than that of the commercial fillers (43.2 ± 8.1%, Restylane; 12.3 ± 5.3%, TheraFill). In addition, immunohistochemical analysis with angiogenesis-related markers such as isolectin and vWF revealed newly formed blood vessels and cellular influx into the composite filler, which were not observed in the other fillers. These results clearly suggest that the HA/COL composite filler is a superior candidate for soft tissue reconstruction. The filler we developed may be a suitable candidate as an injectable dermal filler for tissue augmentation in humans.
Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was conducted for analysis of the purity of umbilicalcord-isolated collagen using 6% polyacrylamide gels and 5% stacking gels at room temperature. Before the electrophoresis, the sample was heated in the presence of sample buffer (70 mM Tris-HCl (pH 6.
The HA/COL composite hydrogels were prepared by blending 2% HA and 2% COL at two different ratios (10:1 and 5:1) using a PT 1200E homogenizer (Kinematica, Luzern, Switzerland) for 3 min. The hydrogels were autoclaved at 121℃ for 20 min for sterilization and were then aliquoted into 1 ml sterilized syringes for experimental use.
대상 데이터
0 ml). For this study, 36 mice were selected to serve as host animals of each injected hydrogel. Two hundred microliters of each prepared HA/COL hydrogel was injected to evenly fill into the backs of the mice.
성능/효과
A 10% or 20% addition of collagen into the HA did not significantly affect its longevity (Fig. 3), while the weight of TheraFill decreased by 88% at 16 weeks.
As shown in Fig. 4, a large number of fibroblast infiltrations were observed in the HA/COL hydrogels, whereas the cells could not be observed in the commercial gels, because the collagen-based filler (TheraFill) has higher viscosity acting as cell barriers, and the HAbased filler (Restylane) has poor cell adhesion due to carboxyl groups in HA. Although a large amount of HA was subcutaneously injected, it moved easily under the skin owing to its hydrophilicity.
The reduced changes in size and weight with time indicate that the corrective effect and aesthetic results can be maintained for a long duration, and the number of injections may therefore be potentially reducible. A 10% or 20% addition of collagen into the HA did not significantly affect its longevity (Fig. 3), while the weight of TheraFill decreased by 88% at 16 weeks.
Consequently, we could expect to prolong dermal correction and to augment tissue post-injection. Although the two HA/COL fillers (10:1 and 5:1) showed better performances than the others, 10:1 (HA/COL) is preferable owing to its longevity in vivo compared with 5:1.
Sixteen weeks after injecting this mixture into mice, in vivo longevity and changes in size and appearance were observed and compared with the effects of commercial dermal fillers. Among the injected fillers, HA/COL (10:1) resulted in a lower weight decrease than Restylane or TheraFill after 16 weeks (Fig. 2).
However, there are still numerous concerns regarding biocompatibility, biosafety, adverse reactions, allergic reactions, inflammation, durability, physical properties, and cost [1]. Approved dermal fillers have been shown to be relatively safe, but varying degrees of resorption make recipients require repeated percutaneous injections to maintain the expected level of collection. Therefore, new dermal filler hydrogels should be able to offer in vivo stability to ensure the longevity of the injectable implant as well as biosafety.
To demonstrate its efficiency, we prepared a material by mixing umbilical-cord-derived COL into HA composite hydrogels. Sixteen weeks after injecting this mixture into mice, in vivo longevity and changes in size and appearance were observed and compared with the effects of commercial dermal fillers. Among the injected fillers, HA/COL (10:1) resulted in a lower weight decrease than Restylane or TheraFill after 16 weeks (Fig.
3A). The 10:1 HA/COL hydrogel showed the lowest weight change rate (24.5 ± 16.9%) among the gels after 16 weeks (Fig. 3B).
2). The reduced changes in size and weight with time indicate that the corrective effect and aesthetic results can be maintained for a long duration, and the number of injections may therefore be potentially reducible. A 10% or 20% addition of collagen into the HA did not significantly affect its longevity (Fig.
후속연구
The filler we developed may be a suitable candidate as an injectable dermal filler for tissue augmentation in humans. Further studies and tests should follow to ensure its safety and efficacy.
Hence we fabricated a prototype of a material that possesses high biocompatibility by mixing human umbilicalcord-derived collagen into cross-linked HA with two ratios (HA:COL = 10:1 and 5:1), because a higher ratio of COL than HA in the hydrogel can result in short durability in vivo, as mentioned above. It can be expected that this composite material will show excellent longevity in clinical applications and good biocompatibility. It has been reported that the combination of HA and collagen often has a positive effect in tissue engineering applications [27].
Approved dermal fillers have been shown to be relatively safe, but varying degrees of resorption make recipients require repeated percutaneous injections to maintain the expected level of collection. Therefore, new dermal filler hydrogels should be able to offer in vivo stability to ensure the longevity of the injectable implant as well as biosafety.
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