UNIVERSIDAD DE GUAYAQUIL

FACULTAD DE INGENIERÍA QUÍMICA

CARRERA INGENIERÍA QUÍMICA

ASIGNATURA
BIOTECNOLOGÍA

CURSO
SEPTIMO SEMETRES “2”

TEMA
DEBER #3

INTEGRANTES
SOLORZANO ALDÁZ KATHERIN
LINO DELVALLE DARWIN
PILOSO PLUAS MARIO
GUIZADO HERRERA DAVID
QUEZADA MUÑOZ PATRICIO

DOCENTE
ING. CECILIA UZCA

CICLO II

2018-2019
 Human ex vivo 3D bone model recapitulates osteocyte response to
metastatic prostate cancer
Abstract

According to a study in the United States, the second leading cause of death in men is due
by the prostate cancer (PCa).

At the moment, there is no type of cure has been found that manages to fight with this
problem, since when establishing this tumor in the bony cavity there is no reversion.

This research carried out in the process shows that 3D culture systems give better results
than using a two-dimensional method because they have a better maturation and in vitro
growth of osteocytes.

By using the Wnt sclerostin inhibitors and adding the dickkopf-1 protein (Dkk-1), a
contrast tendency is demonstrated by decreasing sclerostin and increasing Dkk-1.

By incorporating the Wnt and Dkk-1 proteins, the use of alkaline phosphatase (ALP) is
applied, increasing the presence of mineralization.

Analyzes of 3D culture systems have shown growth and maturation in osteocytes in vitro,
demonstrating an assembly in primary osteocytes of 20-25 μm.

Within the studies carried out, the aggregation of this 3D models of bone tissues with CR
cells that replicating a bone microenvironment to the mast cells is made.

For the execution of this model, the osteocytic FGF23 was applied, giving a
pharmacological finding focused on the PCa.

BACKGROUND
PCa being one of the most prominent causes of death in American citizens, since this is
known as an androgen-driven disease that are the male sex hormones, the morbidity and
mortality of this two is related to be considered as the place or determined point of the PCa
cause. Resulting in metastasis, this being the progression of a tumor disease, becoming
independent of the androgen.

Research is considered important due to tumor cases in poor prognoses and increased
morbidity. Also, it does not have a suitable treatment, to start up the best comprehension of
interactions of the tumor in bone microenvironments.

Through studies carried out towards osteocytes, they have shown that this can influence the
progression of PCa in bone metastasis. The establishment of a point of culture of primary
form in ex vivo in the long term starting from tumor samples of PCa has occurred in a very
difficult way

With the advent of 3D models, the use of the rustic 2D two-dimensional model has been
left aside, given the finding that in human osteocytes small vessels at a micro scale of 20-25
μm can be assembled in small microbeads and cultivated in micro-scale devices.
microfluidic perfusion, replicating the lacunocanalicular structure and functions of human
tissue. Recently the decrease of oxygen in 3D cultures of osteocytic cells was established,
improving the osteocyte phenotype ex vivo. It consists of a formation of a monolayer which
is located between the bone marrow and bone, this is called a critical point in the
interaction of tumor cells with the bone becoming inactive and drug resistant until a
reactivation of the tumor.

MAIN FINDINGS
Through use of Wnt signaling
inhibitors, the osteocytes of 3D
bone tissues are altered, being
exposed to the PCa cell, in this
process the osteocytes are stained in
sections of tissues carrying out the
signaling of both Wnt-scleresotine
and dickkopf-1 (Dkk-1), since the
sclerostin was expressed by the
osteocytes in 3D tissue in the
absence of PCa cells. There was a 6-
fold decrease (p <0.01) in sclerostin,
whereas in Dkk-1 there was a very
significant increase in it, since it
was expressed by osteocytes in 3D
tissues with cell exposure. (3)

Through evaluations between interactions of osteocytes in 2D with PCa cells. When

cultured for 14 days, the osteocytic cells (5 × 10 4) are introduced into PCa cells (2x104)
for a period of 4 more days, such as 3D experimentation. While the expressions of FGF23
and Dkk-1 give low results without differences between crops, as long as crops with
application of ALP give a significant increase (p <0.01) with introductions of PCa cells by
applying 2D, apart from this in 2D media it was analyzed the mineralization without
presenting a significant change.
Through these studies, the CR cells in the PCa give the facility to imitate the bone, by
showing phenotypes, being the set of genes of an organism and genotypes that is the
representative form of denoting a cancer cell. These studies are given to verify genetic
changes observed in bone tissues, being cultured for 4 days given these data the PCa cells
didn’t express osteogenic markers, on the other hand, the SOST is given at low levels,
without changes in culture media 1:1.
DISCUSSION

The implementation of biologically relevant tissue 3D models is due to the lack of

recapitulation of microenvironmental factors as critical points of development and
medication evaluations. Given the present study is given as a source of development
towards metastatic PCa investigations with the implementation of osteocytes of ex vivo
characterizations, giving the application of FGF23 as an emerging measure in cancer and
other cases, such model gives as a new source of applications replacing use of animals,
reducing cellular limitations of PCa.

The review of the study results in three types of analysis in models such as: assembly to 3D
devices of bone metastasis, assembly assisted by 3D matrix of bone metastasis, direct 3D
bone tumor, in contrast to 23 systems examined obtaining unique characteristics: primary
osteocytes human, hypoxic conditions, integration with CR cells instead of using
conventional cell lines. The need to use human osteoblasts is recognized, improving the
biometric nature in human bone tissues, this integration is useful in dynamic intercellular
communications in co-culture models, such culture results in the recapitulation of bone
microenvironment and lines PC3 replacing PCa.

It has been demonstrated, FGF23 promotes PCa progressions increasing the formation of
bone metastasis, while PCa cells are capable of secreting and expressing receptors,
increasing levels of FGF23, in turn giving rise to PCA cells, when he found
immunofluorescence staining in presence, validating the research with genetic expressions
and confirming the significant increase of FGF23 in 3D bone tissues cultured in PCa cells.

Given relevance of the study, it’s shown that the 3D model has an exorbitant power, in
interactions with disseminated tumor cells.
 Deber 3: Rúbrica revision
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