Collagen Type 0

Biocompatibility properties of Collagen Type 0 (in vitro)

In this in vitro study the effect of Collagen Type 0 on differentiated THP-1 cells, a type of cell associated with wound development, was evaluated. A previous study reported that collagen extracted from other jellyfish species (Nemopilema nomurai) may stimulate the secretion of pro-inflammatory cytokines (TNF-α / IL-6) (Ref- Putra A.B.D.N et. al., 2014). We therefore wanted to see if Collagen Type 0 from R. pulmo shows a similar or different effect.

Data from our study (Fig. 1) demonstrated that Collagen Type 0 did not induce secretion of NF-kB regulated cytokines TNF-α and IL-8 so does not mediate inflammation through the NF-kB pathway in dTHP-1 cells.

Inflammatory cytokine secretion

The secretion of TNF-α, together with other cytokines associated with inflammation (IL-8, IL-1β and IL-18), were evaluated in the differentiated THP-1 cells alone or when Collagen Type 0 was present. It was found that Collagen Type 0 did not induce significant changes in the secretion of pro-inflammatory cytokines (Fig. 1). Furthermore, the differentiated THP-1 cells (both alone and when Collagen Type 0 was present) were challenged with lipopolysaccharide (LPS). LPS is a major component of gram negative bacteria and is known to trigger an inflammatory immune response. Introduction of LPS led to a marked inflammatory response, but the presence of Collagen Type 0 did not alter this in any way (Fig. 2).

Key takeaway: Collagen Type 0 shows excellent biocompatibility and does not act in a proinflammatory (via Nf-KB or NLRP3 Inflammasome pathways) way in the presence of macrophages.

Are anti-inflammatory properties of Collagen Type 0 also seen in vivo?

A 2020 study by Flaig I. et al. examined the macrophage response to Collagen Type 0 implanted subcutaneously in Wistar rats. A control group were implanted with porcine pericardium collagen (PPC).

Total macrophage numbers

There were significantly lower numbers of total macrophages in response to Collagen Type 0 compared to PPC at both days 10 and 30 (p<0.001) (Fig. 1). Total macrophage numbers did not significantly decrease between day 10 and day 30 with PPC, while there was a significant two-fold reduction with Collagen Type 0 (p<0.05)*.

M1 and M2 macrophage balance

Looking at the balance between M1 and M2 macrophages, there were significantly more M2 macrophages than M1 at day 10 with Collagen Type 0 (p<0.05) (Fig. 2.1, Fig. 2.2). By day 30, total macrophages had decreased and there was no significant difference between the two types present in the wound bed. With PPC, there was only a shift to predominantly M2 macrophages after 30 days*. Typically, in a wound, most healing occurs by day 30 and the body will go into an equilibrium state.

*Results from an unpublished sub-analysis of the data

Proteomics

Bowen A. et al. (2022) compared the tissue reaction of Collagen Type 0 to hyaluronic acid and human cadaveric dermis using proteomic profiles of rabbits following a bulk filler injection of Collagen Type 0 into vocal folds. Data showed that Collagen Type 0 uniquely upregulates pathways related to ECM/ collagen formation suggesting tissue restorative properties and downregulates pathways related to inflammation, including T1 inflammation an indication of very low foreign body reaction (Fig. 3).

Collagen Type 0 was also found to promote energy production through beta acid oxidation, potentially triggering pro-regenerative responses.

Impact on healing of a cranial wound

In this in vivo study it was investigated whether the anti-inflammatory properties of Collagen Type 0 can impact the healing of a cranial wound in rats.

Bone formation

When examining bone formation, there was >45% bone growth in the Collagen Type 0 group with a significantly lower percentage (9%) in the group with no collagen implanted (Fig. 1).

Wound healing

In this in vivo study, we looked at whether Collagen Type 0 can close and heal full-thickness excisional wounds where there is impaired healing. A diabetic mouse model was used to simulate impaired wound healing.

Wounds treated with Collagen Type 0 reached 80% closure faster than those not treated. Collagen Type 0 resulted in superior uniformity and complete wound closure (Fig. 2.1, Fig. 2.2).

Bulk filling

Using an injectable laryngoplasty model, Collagen Type 0 was evaluated as an injectable bulk filler material. MRI data demonstrates that Collagen Type 0 outperforms standard of care hyaluronic acid and human collagen type I and provides improved medialisation as well as increased glottal benefit with minimal tissue pathology (Fig. 3.1, 3.2) (San Marina. S., et al. Laryngoscope 2021).

Collagen Type 0 Characteristics

Collagen Type 0 is a fibrillogenic, triple helical collagen that is produced by Jellagen with batch-to-batch consistency and key characteristics as described below.