81
of CS from cartilage of the lesser spotted dogfi sh ( Scyliorhinus canicula ), was reported
by Gargiulo et al. 2009 . This representative the Scryliohinidae family of the
Carcharhiniformes order (Delabre et al. 1998 ) is broad distributed around the coast of
the Mediterranean Sea, however, has no marketability. Unfortunately, regularly a large
number of specimens die because they are erroneously captured during fi shing
expeditions. “The dead animals are usually thrown back to the sea, wasting the
opportunity to isolate chondroitin sulfate from their tissues and use it for commercial
preparations” (Gargiulo et al. 2009 ).
Collagen is a well-known as well as well studied biological material of proteinaceous origin (see also Chap. 8 in this work). Here, only one example of it use as a
scaffold. Recently, Sangsen and co-workers ( 2012 ) reported about development of
composite material that consist of fi sh collagen and plant pectin. They used the skins of
brown banded bamboo sharks ( Chiloscyllium punctatum ) and citrus fruits for isolation
of collagen and pectin, respectively. Crosslinking of these novel scaffolds was performed by chemical crosslinking of shark’s collagen using carbodiimide and by pregelation of pectin using calcium sulphate. The materials properties of the hybrid scaffolds
obtained were as follow: average pore size of 134.53 ± 52.44 μm for the interconnecting pores; the hardness of 0.591 ± 0.135 N, and a springiness of 0.958 ± 0.022 (Sangsen
et al. 2012 ). The scaffolds were biocompatible as shown by an in vitro cytotoxicity test
using C 2 C1 2 myoblast cells. The authors propose these fi sh collagen-pectin scaffolds as
composite materials suitable for applications in tissue engineering.
However, is it possible to cultivate cartilage cells from marine vertebrates and use
them as model systems, or even directly for tissue engineering? To my best knowledge, there are only few papers related to this topic. For example, as previously
reported by Langille and Hall ( 1988 ), “cartilage from larval (ammocoetes) and adult
(prespawning upstream migrant) lamprey was successfully maintained both when
cultured in vitro and when grafted in vivo on the chorioallantoic membrane of host
chick embryos,” (Langille and Hall 1988 ). Moreover, it is possible to cultivate teeth
from an adult lamprey under in vitro conditions. Histological investigations showed
with strong evidence the cellular and structural integrity of both the cultured and
grafted cartilages. Also ultrastructural analysis of chondrocytes using TEM confi rmed the viability of the cartilage. Additionally, in vitro incorporation of radioactive
sulfur into the matrix has shown metabolic activity of this tissue. Thus, “teeth cultured in L15-supplemented media for up to 14 days at either 15 or 20 °C retained
their structural and cellular integrity as observed histologically, with no apparent cell
outgrowth” (Langille and Hall 1988 ). There are no doubts that with the successful
culture of the selected lamprey’s tissues, their biochemistry, physiology and development, are potentially of great importance in better understanding of early vertebrate
evolution and opens new ways in tissue engineering of marine cartilages.
Moreover, the phenomenon of in vitro mineralization of both lamprey teeth and
cartilage in culture is one of the principal questions. The capability of lamprey cartilage to calcify in vitro under selected ionic conditions has been also reported
(Langille and Hall 1985 ). Numerous experiments with respect to obtaining of
hydroxyapatite have been carried out using adult and larval lamprey cartilage, normally unmineralized. The results of these 12 days- long experiments, which were
carried out at 20, 30 or 37 °C are as follow:
2.1 From Non-mineralized to Mineralized Cartilage
of CS from cartilage of the lesser spotted dogfi sh ( Scyliorhinus canicula ), was reported
by Gargiulo et al. 2009 . This representative the Scryliohinidae family of the
Carcharhiniformes order (Delabre et al. 1998 ) is broad distributed around the coast of
the Mediterranean Sea, however, has no marketability. Unfortunately, regularly a large
number of specimens die because they are erroneously captured during fi shing
expeditions. “The dead animals are usually thrown back to the sea, wasting the
opportunity to isolate chondroitin sulfate from their tissues and use it for commercial
preparations” (Gargiulo et al. 2009 ).
Collagen is a well-known as well as well studied biological material of proteinaceous origin (see also Chap. 8 in this work). Here, only one example of it use as a
scaffold. Recently, Sangsen and co-workers ( 2012 ) reported about development of
composite material that consist of fi sh collagen and plant pectin. They used the skins of
brown banded bamboo sharks ( Chiloscyllium punctatum ) and citrus fruits for isolation
of collagen and pectin, respectively. Crosslinking of these novel scaffolds was performed by chemical crosslinking of shark’s collagen using carbodiimide and by pregelation of pectin using calcium sulphate. The materials properties of the hybrid scaffolds
obtained were as follow: average pore size of 134.53 ± 52.44 μm for the interconnecting pores; the hardness of 0.591 ± 0.135 N, and a springiness of 0.958 ± 0.022 (Sangsen
et al. 2012 ). The scaffolds were biocompatible as shown by an in vitro cytotoxicity test
using C 2 C1 2 myoblast cells. The authors propose these fi sh collagen-pectin scaffolds as
composite materials suitable for applications in tissue engineering.
However, is it possible to cultivate cartilage cells from marine vertebrates and use
them as model systems, or even directly for tissue engineering? To my best knowledge, there are only few papers related to this topic. For example, as previously
reported by Langille and Hall ( 1988 ), “cartilage from larval (ammocoetes) and adult
(prespawning upstream migrant) lamprey was successfully maintained both when
cultured in vitro and when grafted in vivo on the chorioallantoic membrane of host
chick embryos,” (Langille and Hall 1988 ). Moreover, it is possible to cultivate teeth
from an adult lamprey under in vitro conditions. Histological investigations showed
with strong evidence the cellular and structural integrity of both the cultured and
grafted cartilages. Also ultrastructural analysis of chondrocytes using TEM confi rmed the viability of the cartilage. Additionally, in vitro incorporation of radioactive
sulfur into the matrix has shown metabolic activity of this tissue. Thus, “teeth cultured in L15-supplemented media for up to 14 days at either 15 or 20 °C retained
their structural and cellular integrity as observed histologically, with no apparent cell
outgrowth” (Langille and Hall 1988 ). There are no doubts that with the successful
culture of the selected lamprey’s tissues, their biochemistry, physiology and development, are potentially of great importance in better understanding of early vertebrate
evolution and opens new ways in tissue engineering of marine cartilages.
Moreover, the phenomenon of in vitro mineralization of both lamprey teeth and
cartilage in culture is one of the principal questions. The capability of lamprey cartilage to calcify in vitro under selected ionic conditions has been also reported
(Langille and Hall 1985 ). Numerous experiments with respect to obtaining of
hydroxyapatite have been carried out using adult and larval lamprey cartilage, normally unmineralized. The results of these 12 days- long experiments, which were
carried out at 20, 30 or 37 °C are as follow:
2.1 From Non-mineralized to Mineralized Cartilage