# Three-Dimensional Human Liver Micro Organoids and Bone Co-Culture Mimics Alcohol-Induced BMP Dysregulation and Bone Remodeling Defects

## TL;DR

This study created a 3D human liver-bone model that mimics alcohol-related liver and bone damage, showing how alcohol disrupts bone proteins and weakens bone health.

## Contribution

The study introduces a 3D human liver-bone co-culture model that replicates alcohol-induced hepatic osteodystrophy and BMP dysregulation.

## Key findings

- Chronic alcohol exposure activates CYP2E1 and causes liver fibrosis and EMT.
- Alcohol reduces BMP2 and increases BMP13, leading to impaired osteoblast function and chondrogenic shifts in bone progenitors.
- The model reflects clinical features of chronic liver disease, enabling BMP-targeted therapy screening.

## Abstract

What are the main findings?
The present study established a long-term 3D human liver–bone co-culture model mimicking alcohol-induced hepatic osteodystrophy (HOD) with fibrogenic liver changes and bone defects.Chronic 50 mM alcohol triggers hepatic CYP2E1 activation, EMT/fibrosis, BMP imbalance (↓BMP2, ↑BMP13), reduced osteoblast mineralization, and chondrogenic shift in bone progenitors.

The present study established a long-term 3D human liver–bone co-culture model mimicking alcohol-induced hepatic osteodystrophy (HOD) with fibrogenic liver changes and bone defects.

Chronic 50 mM alcohol triggers hepatic CYP2E1 activation, EMT/fibrosis, BMP imbalance (↓BMP2, ↑BMP13), reduced osteoblast mineralization, and chondrogenic shift in bone progenitors.

What are the implications of the main findings?
The present study provides an organoid-based platform for studying the liver–bone axis and BMP dysregulation in HOD, surpassing monocultures or animal models.The present study enables the screening of BMP-targeted therapies to restore bone homeostasis in alcohol-related chronic liver disease.

The present study provides an organoid-based platform for studying the liver–bone axis and BMP dysregulation in HOD, surpassing monocultures or animal models.

The present study enables the screening of BMP-targeted therapies to restore bone homeostasis in alcohol-related chronic liver disease.

Hepatic osteodystrophy (HOD) is a frequent complication of chronic liver disease, marked by impaired osteogenesis and elevated fracture risk, particularly under sustained alcohol exposure. Bone morphogenetic proteins (BMPs), which play a crucial role in maintaining bone homeostasis, are dysregulated in alcoholic liver disease. Specifically, decreased BMP2 and increased BMP13 have been linked to impaired osteogenesis and cartilage-like shifts in bone progenitors. A human in vitro system that recapitulates this hepatic BMP imbalance is needed to dissect mechanisms and identify targets. To address this, we established a long-term human three-dimensional liver–bone co-culture model that integrates hepatocytes (HepaRG), hepatic stellate cells (LX-2), and human umbilical vein endothelial cells (HUVECs) with bone scaffolds seeded with osteoblast precursors (SCP-1) and osteoclast precursors (THP-1). This study aimed to characterize the effects of chronic 50 mM alcohol exposure on hepatic fibrogenic activation and BMP ligand secretion, and to investigate the associated BMP-responsive signaling involved in bone cell lineage differentiation and functional activity. The results demonstrated alcohol-induced hepatic CYP2E1 activation and fibrogenic remodeling with EMT signatures, as well as a decrease in BMP2 and an increase in BMP13, without affecting BMP9. Liver-derived factors activated both canonical and non-canonical BMP signaling in bone progenitors, reduced osteoblast activity and mineralization, preserved osteoclast TRAP activity, and shifted the lineage toward chondrogenesis (SOX9↑, RUNX2↓). Notably, this BMP profile and skeletal phenotype reflect clinical observations in chronic liver disease, indicating that the model recapitulates key in vivo pathological features. This human liver micro-organoid co-culture reproduces alcohol-induced hepatic BMP dysregulation and downstream bone defects, offering an organoid-centric, microengineered platform for mechanistic studies and BMP-targeted therapeutic screening in HOD.